Abstract: The invention is directed to the use of RNA or DNA polymerases having 3'-intrinsic editing activity (3'-IEA) in the presence or absence of a deactivating agent to remove a protecting group from the 3'-position of oligo- polyribo- or deoxyribonnucleotides. The use is connected with the incorporation of dNTPs into DNA templates in order to determine the concentration and/or sequence of said templates. In particular the use is concerned with an improved non gel-based sequencing method.

Abstract: Methods of synthesizing polynucleotides are disclosed involving the simultaneous ligation of a set of oligomer 5'-phosphates onto a template-bound primer. The set of these oligomers can be preselected to contain oligomers which are complementary to the template strand or the oligomers can be supplied as a library and allowed to self select. The synthesis by ligation can proceed unidirectionally or bidirectionally from the primer and can be used to synthesize both strands simultaneously by the use of two primers. The ligation is preferably performed with a ligase enzyme. The methods of synthesis are useful in a variety of applications, including cloning, amplification, labeling, diagnostic assays, mutation analysis and screening, gene expression monitoring and sequence analysis.

Abstract: The present invention relates to a nucleic acid sequence isolated from C. coli which hybridizes specifically with the genomic nucleic acid of strains belonging to the species C. coli and which does not hybridize under the usual conditions with the nucleic acids of campylobacteria which do not belong to this species, nor with the genomic nucleic acids of bacteria related to the genus Campylobacter.

Abstract: The present invention provides a method for preparing HSV vectors. The method comprises co-transfecting a source vector and a mutating cassette together into a population of appropriate host cells, such that homologous recombination occurs between the mutating cassette and the source vector whereby the mutating cassette replaces a region of the HSV genome. The mutating cassette has a unique restriction site not present in the sequence of the vector. The method further comprises plaquing the co-transfected host cells, selecting plaques in which recombination has occurred between the source vector and the mutating cassette, and isolating the viral DNA from the plaques. The isolated viral DNA is digested with a restriction endonuclease appropriate for cleaving the viral DNA at the unique restriction site within the mutating cassette to produce two viral polynucleotides. Following purification, the two viral polynucleotides can be ligated to form an HSV vector comprising the two viral polynucleotides.

Abstract: A method of amplifying a nucleotide sequence complementary to an mRNA template derived from genomic DNA. The method involves the following steps. A sample mixture containing an mRNA template and corresponding genomic DNA is provided, the genomic DNA including at least two exons separated by at least one intron. A pair of intron-blockers are introduced into the mixture, the intron-blockers comprising a sequence of intron-specific oligonucleotides modified to prevent nucleotide extension in conditions promoting polymerase chain reaction. A primer pair promoting amplification of cDNA derived from the mRNA template is introduced into the mixture and then reverse transcription polymerase chain reaction is carried out to amplify cDNA. Detection of the cDNA is proof of the existence of mRNA in the sample, and thus proof of expression of the corresponding gene. The method avoids false positives caused by amplification of genomic DNA as well as cDNA based on an mRNA template.

Abstract: The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof. The present invention further relates to methods and devices for the separation of nucleic acid molecules based on charge. The present invention also provides methods for the detection of non-target cleavage products via the formation of a complete and activated protein binding region. The invention further provides sensitive and specific methods for the detection of human cytomegalovirus nucleic acid in a sample.

Abstract: A method of amplifying a nucleotide sequence complementary to an mRNA template derived from genomic DNA. The method involves the following steps. A sample mixture containing an mRNA template and corresponding genomic DNA is provided, the genomic DNA including at least two exons separated by at least one intron. A pair of intron-blockers are introduced into the mixture, the intron-blockers comprising a sequence of intron-specific oligonucleotides modified to prevent nucleotide extension in conditions promoting polymerase chain reaction. A primer pair promoting amplification of cDNA derived from the mRNA template is introduced into the mixture and then reverse transcription polymerase chain reaction is carried out to amplify cDNA. Detection of the cDNA is proof of the existence of mRNA in the sample, and thus proof of expression of the corresponding gene. The method avoids false positives caused by amplification of genomic DNA as well as cDNA based on an mRNA template.

Abstract: Activation of NF-AT-dependent transcription, including agents which interfere with the production, modification of the nuclear or cytoplasmic subunits, or the nuclear import of the cytoplasmic subunits. In particular, screening tests for novel immunosuppressants are provided based upon the ability of NF-AT to activate transcription.

Abstract: In a method for amplifying the specific nucleic acid sequence, a highly specific amplification which has low possibility of non-specific hybridization can be carried out. Highly stable reagents, the activities of which do not decrease in case of supply and storage, are also provided. Thermostable enzymes are used as RNA dependent DNA polymerase, DNA dependent DNA polymerase, DNA dependent RNA polymerase and ribonuclease H, which are required for the amplification system based on replicated RNA. Especially, it is preferable that a thermostable enzyme derived from Thermus thermophilus which has RNA dependent DNA polymerase activity, DNA dependent DNA polymerase activity and ribonuclease H activity, and thermostable DNA dependent RNA polymerase are used together. By this method, inactivation of enzymes are prevented by using thermostable enzymes, and the amplification can be carried out without sequential addition of enzymes.

Abstract: A method for enhancing the detection of prostate specific antigen in a biological sample comprising (a) extracting mRNA from the sample: (b) contacting the mRNA from step (a) with reverse transcriptase under conditions allowing for the production of cDNA; (c) contacting the cDNA from step (b) with a pair of reverse transcriptase polymerase chain reaction primers capable of specifically hybridizing with DNA encoding prostate specific antigen wherein one such primer is an oligonucleotide of 12 to 30 nucleotides in length and comprises the sequence 5-CACCCATCCTA-3' and wherein the second such primer is an oligonucleotide of 12 to 30 nucleotides in length and comprises the sequence 5'-TCCAGCCACGAC-3'; and wherein at least one of the primers is covalently linked to a modified digoxigenin molecule and under conditions allowing for extension of the primers; and (d) detecting the resulting amplified DNA.

Abstract: Disclosed are diagnostic techniques for the detection of human prostate disease. The invention relates particularly to probes and methods for evaluating the presence of RNA species that are differentially expressed in metastatic prostate cancer compared to normal human prostate, benign prostatic hyperplasia, and non-metastatic prostate cancer. The invention also relates to probes and methods for evaluating the presence of RNA species that are differentially expressed in the peripheral blood of individuals with the disease state compared to normal healthy individuals. Described are methods of therapeutic use for genes identified as differentially expressed in metastatic prostate cancer, and means for screening pharmaceuticals effective in treatment of prostate cancer.

Abstract: The present invention provides a dry solid medium for storing a sample of a genetic material, including RNA and DNA, in a form suitable for subsequent analysis. The dry solid medium of the invention includes a dry solid matrix and a composition including a protein denaturing agent, and a chelating agent. The dry solid medium can further include a component which functions in subsequent analysis of the genetic material using, for example, PCR, reverse transcriptase initiated PCR, LCR, RFLP, or genetic hybridization. A component for subsequent analysis includes, for example, a nucleotide sequences such as a primer, DNA or RNA probe, and a target sequence stabilizer. The dry solid medium can also include a retaining agent to enhance retention of a component for subsequent analysis. The invention also provides methods for using the dry solid medium of the invention. The dry solid media and methods of use thereof are particularly suited for use in automated systems.

Abstract: A system for isolating mRNAs as cDNAs employs a polymerase amplification method using at least two oligodeoxynucleotide primers. In one approach, the first primer contains sequence capable of hybridizing to a site immediately upstream of the first A ribonucleotide of the mRNA's polyA tail and the second primer contains arbitrary sequence. In another approach, the first primer contains sequence capable of hybridizing to a site including the mRNA's polyA signal sequence and the second primer contains arbitrary sequence. In another approach, the first primer contains arbitrary sequence and the second primer contains sequence capable of hybridizing to a site including the Kozak sequence. In another approach, the first primer contains a sequence that is substantially complementary to the sequence of a mRNA having a known sequence and the second primer contains arbitrary sequence.

Abstract: Asn-linked glycoprotein having antitumor activity, with molecular weight of 32-84 kDa, its producing strain, production and use, adsorbs to Con. A. The process for its production comprises culturing a microorganism belonging to genus Aspergillus and isolating from the culture medium Asn-linked glycoprotein having a molecular weight of 32-84 kDa which adsorbs to Con. A, or a mixture thereof. The substance having antitumor activity is effective for treatment of solid tumors, ascites tumors, multiple cytoma and oval tumors and for the suppression of tumors.

Abstract: Novel polynucleotides and the proteins encoded thereby are disclosed.

Abstract: A method is provided for genotyping a target sequence at at least two allelic sites by a 5' nuclease amplification reaction. In one embodiment, the method includes performing a nucleic acid amplification on a target sequence having at least two different allelic sites using a nucleic acid polymerase having 5'.fwdarw.

Abstract: The invention provides a method for diagnosis of genetic abnormalities or other genetic conditions which can be readily automated. The method is used to determine the presence or absence of a target sequence in a sample of denatured nucleic acid and entails hybridizing the sample with a probe complementary to a diagnostic portion of the target sequence (the diagnostic probe), and with a probe complementary to a nucleotide sequence contiguous with the diagnostic portion (the contiguous probe), under conditions wherein the diagnostic probe remains bound substantially only to the sample nucleic acid containing the target sequence. The diagnostic probe and contiguous probe are then covalently attached to yield a target probe which is complementary to the target sequence, and the probes which are not attached are removed.

Abstract: Novel polynucleotides and the proteins encoded thereby are disclosed.

Abstract: Provided herein are methods for detecting multiple target nucleic acid sequences in a test sample. Also provided is a hybridization platform useful for detecting multiple target sequences in a test sample. The hybridization platform comprises a solid support material having a defined pattern of capture probes immobilized thereon.

Abstract: The present invention relates to a human lactoferrin cDNA obtained from human breast tissue and the protein encoded therefrom. The present invention further relates to methods for detecting malignancy arising from tissues that normally secrete lactoferrin using the cDNA gene probe of the present invention. Another aspect of the present invention relates to the promotor region that regulates the human lactoferrin gene.

Abstract: DNA ladders are constructed by partial restriction digestion of plasmids with the following design properties: (a) the plasmid contains five or more sites of a restriction endonuclease; (b) the length between adjacent sites is an integer multiple of a minimal length, said minimal length is the length between the closest adjacent sites; (c) partial restriction endonuclease digestion of the plasmid generates a DNA ladder with at least five differently sized DNA fragments; and (d) said DNA ladder has the properties that (1) all DNA fragments of the ladder have lengths which are integer multiples of the minimal length; (2) the smallest DNA fragment length is the minimal length, (3) the largest DNA fragment is the length of the entire plasmid; and (4) the ladder has a DNA fragment at every integer multiple of the minimal length ranging from the minimal length to the plasmid length. This plasmid design is especially useful for making small fragment DNA ladders, such as 100 and 200 base pair ladders.

Abstract: A gene is provided which is present in the deletion region of a chromosome common in lung cancer, hepatocellular carcinoma and colorectal cancer and encodes a novel protein, a protein encoded by the gene (PRLTS protein), and a method of discriminating tumor cells.

Abstract: The invention relates to a method of synthesizing polydeoxyribonucleotides which comprises causing a thermostable deoxyribunucleotide polymerase to act on deoxyribonucleotides without using any template and any primers, to the thus-obtained polydeoxyribonucleotides, to a method of screening cDNA libraries using the same as probes, to a method comprising joining the DNA resulting from the above method of synthesis to the 3'--OH terminus of a double-stranded DNA containing a gene derived from cells of a living organism, followed by transfection of such cells with the product of joining and culture of the cells, to thereby cause insertion of the product into chromosomal DNA and production of the protein encoded by the gene, to a method of synthesizing chromosomal DNA and to a method of synthesizing chromosomes.

Abstract: The nucleic acid amplification procedures of the present invention provide methods for amplification of a nucleic acid comprised of a first strand, comprising (a) using the first strand to generate copies of a second strand at a first location, (b) moving the copies of the second strand to a second location, and (c) using the copies of the second strand to generate copies of at least a portion of the first strand. Target nucleic acids used in the context of the present method include RNA or DNA, either single stranded or double stranded, using primer extension or joining-type protocols. Embodiments are set forth for automated forms of the claimed procedures.

Abstract: In order to sequence nucleic acids one produces a mixture of labelled nucleic acid fragments of different length, using nucleic acid fragments which are labelled by incorporation of at least one deoxyribonucleoside triphosphate with a non-radioactive labelling group, separates the labelled nucleic acid fragments according to size and determines the nucleic acid sequence by means of the labelling of the individual fragments.

Abstract: Telomerase activity in a sample can be measured using a two reaction protocol. The first reaction involves the formation of telomerase substrate extension products from a telomerase substrate. The second reaction involves replication of the telomerase substrate extension products and/or amplification of signal generated by a bound probe. The presence of telomerase activity in a human somatic tissue or cell sample is positively correlated with the presence of cancer and can be used to diagnose a disease or other conditions of medical interest, as well as the course of disease progression or remission in a patient.

Abstract: An oligonucleotide is intended to be used as a promoter non-template strand in the transcription of a sequence of a nucleotide target in the presence of a phage RNA polymerase. The phage RNA polymerase has specific natural promoters containing a consensus sequence from at least position -17 to position -1. The oligonucleotide contains a core sequence flanked at at least one of its ends by a nucleotide sequence capable of hybridization with a sequence of the target. The core sequence contains a sequence of 6 to 9 consecutive nucleotides from the region -12 to -4 of the non-template strand of the specific promoter, or a sufficiently homologous sequence to enable the functionality of the RNA polymerase to be retained.

Abstract: A method, composition and kit for amplifying a target nucleic acid sequence under conditions of substantially constant temperature, ionic strength, and pH and using only a single promoter-primer. To effect the amplification, a supply of a single promoter-primer having a promoter and a primer complementary to the 3'-end of the target sequence, and a reverse transcriptase and an RNA polymerase are provided to a mixture including the target sequence; the amplification proceeds accordingly. The invention is useful for generating copies of a nucleic acid target sequence for purposes that include assays to quantitate specific nucleic acid sequences in clinical, environmental, forensic and similar samples, cloning and generating probes.

Abstract: The invention relates to methods for determining sequence information in polynucleotides by combining the recent disparate technologies of mass spectrometry and polynucleotide hybridization, amplification, extension and/or ligation techniques. Broadly, in a first step, the method for determining sequence information in a sample polynucleotide includes hybridizing with a sample nucleotide one or a mixture of oligonucleotide probes having a nucleotide sequence complementary to a portion of the sample polynucleotide, thereby forming a complex. Then, in a second step, the complex is contacted with at least a member selected from the group consisting of nucleosides, dideoxynucleosides, polymerases, nucleases, transcriptases, ligases and restriction enzymes to alter at least a subset of said oligonucleotide probes.

Abstract: Disclosed are diagnostic techniques for the detection of human prostate cancer. Genetic probes and methods useful in monitoring the progression and diagnosis of prostate cancer are described. The invention relates particularly to probes and methods for evaluating the presence of RNA species that are differentially expressed in prostate cancer compared to normal human prostate or benign prostatic hyperplasia.

Abstract: The subject invention is directed to a method for determining the integrity of nucleic acid isolated from a specimen of eukaryotic origin comprising subjecting the specimen to nucleic acid isolation followed by amplification using a pair of oligonucleotide primers in a manner known per se, said primers being capable of amplifying nucleic acid specific for RNA encoding a product of a housekeeping gene, said RNA being low abundance RNA, said gene being present in all cell types of a eukaryotic species, and said method further comprising determining whether amplification of the nucleic acid specific for the RNA has occurred. In particular, the method comprises amplifying mRNA. Optionally, the method can also comprise quantifying the amplification product. As this method is of particular relevance for diagnosis of diseases in humans, the eukaryotic species is preferably a human being.

Abstract: A method of detecting a nucleic acid (e.g., DNA, RNA) that contains at least one preselected base (e.g., adenine, guanine, 6-mercaptoguanine, 8-oxo-guanine, and 8-oxo-adenine) comprises (a) reacting the nucleic acid with a transition metal complex capable of oxidizing the preselected base in an oxidation-reduction reaction; (b) detecting the oxidation-reduction reaction; and (c) determining the presence or absence of the nucleic acid from the detected oxidation-reduction reaction at the preselected base. The method may be used in a variety of applications, including DNA sequencing, diagnostic assays, and quantitative analysis.

Abstract: Described is a support material that can simultaneously bind both genotypic and phenotypic substances. The respective areas, which can have surface-modifying matter such as anionic exchangers and/or affinity ligands, simultaneously bind, for example, nucleic acids and proteins or peptides. The support materials described can be used in processes for evolutive optimization of biopolymers, wherein genotype and phenotype can be bound at the same time so as to furnish them for further analysis.

Abstract: The present invention pertains to an in vitro method for assaying for an inhibitor of the catalytic Group I self-splicing intron reaction in the nuclear rRNA genes of Pheumocystis carinii which comprises the steps of (a) providing a DNA template containing the intron (I) from the 26S rRNA gene in Pneumocystis carinii and a portion of the 5' and 3' flanking exons (E1 and E2, respectively) between nucleotides 1963 and 2267 of 26S rRNA (660 nucleotides of amplified rRNA gene including the group I intron); (b) preparing an RNA precursor by transcription of the DNA template in the presence of labeled nucleoside triphosphates to produce a labeled RNA precursor (E1-I-E2); (c) purifying the RNA precursor; (d) incubating the RNA precursor and the inhibitor in the presence of guanosine triphosphate and magnesium ions; and (e) determining the degree of inhibition by the inhibitor on the intron splicing reaction in the RNA precursor by measuring the amount of labeled splicing intermediates and splicing products.

Abstract: A method for synthesizing a nucleic acid molecule comprising at least one non-canonical nucleoside triphosphate using a mutant polymerase having a reduced discrimination between canonical and non-canonical substrates is disclosed. The method comprises incubating a template nucleic acid in a reaction mixture comprising the mutant nucleic acid polymerase and the appropriate canonical and non-canonical nucleoside triphosphates which are desired substrates for the mutant nucleic acid polymerase. The present invention is also a method of determining the sequence of a nucleic acid molecule using the mutant polymerase to create a nucleic acid molecule comprising at least one non-canonical nucleoside triphosphate.

Abstract: The design of new ribozymes capable of self-catalyzed trans-splicing which are based upon the catalytic core of a Group I intron are described. Using this design, it is possible to construct ribozymes capable of efficiently splicing a new 3' exon sequence into any chosen target RNA sequence in a highly precise manner. A method of cell ablation is also described that provides a toxic product to a host cell in vivo in a targetted, regulated manner utilizing novel trans-splicing ribozymes of the invention. Inactive pro-ribozyme forms are also described.

Abstract: The invention provided for a method of transcriptionally modulating the expression of a gene encoding a protein of interest associated with treatment of one or more symptoms of a cardiovascular disease. Further provided is a method of determining whether a molecule not previously known to be a modulator of protein biosynthesis is capable of directly and specifically transcriptionally modulating the expression of a gene encoding a protein of interest associated with treatment of one or more symptoms of a cardiovascular disease. Screening methods, including methods of essentially simultaneously screening molecules to determine whether the molecules are capable of directly and specifically transcriptionally modulating one or more genes encoding proteins of interest associated with treatment of one or more symptoms of a cardiovascular disease, are also provided.

Abstract: The present invention relates, in general to protein that is a seed storage protein having high nutritional value. In particular, the invention relates to the protein AmA1 and to a DNA sequence encoding same. The invention further relates to a recombinant molecule comprising the AmA1 encoding sequence and to a host cell transformed therewith. In addition, the invention relates to a method for producing transgenic plants with high nutritionally rich amino acids.

Abstract: Disclosed is the family of genes responsible for the neurodegenerative diseases, particularly Amyotrophic Lateral Sclerosis. Methods and compounds for the diagnosis, prevention, and therapy of the disease are also disclosed.

Abstract: Method and compositions are provided for the determination of telomere length and telomerase activity, as well as the ability to inhibit telomerase activity in the treatment of proliferative diseases. Particularly, primers are elongated under conditions which minimize interference from other genomic sequences, so as to obtain accurate determinations of telomeric length or telomerase activity. In addition, compositions are provided for intracellular inhibition of telomerase activity.

Abstract: The present invention relates to genetic mutations in mitochondrial genes that segregate with diabetes mellitus. The invention provides methods for detecting such mutations, as a diagnostic for diabetes mellitus, either before or after the onset on clinical symptoms. Examples of specific mutations in the mitochondrial ATP synthase 8/6 gene and tRNA lysine gene are given. The invention also provides treatments for dysfunctions due to mitochondrial genes that segregate with diabetes mellitus. Cybrid cell lines are described which are useful as model systems for the study of the mitochondrial metabolic disorders that are associated with diabetes mellitus, and for identifying therapeutic compounds and treatments for this disease.

Abstract: A method by which a nucleotide sequence, specifically a CAG triplet repeat shown to be expanded in individuals with Machado-Joseph Disease can be identified in a sample obtainable from an individual. The present methods can be used to identify individuals in whom the CAT triplet repeat is expanded, including methods useful to identify the protein encoded by the Machado-Joseph Disease gene.

Abstract: The present invention provides a novel method for detecting the presence, or absence, or reduced level of an mRNA transcript of a novel tumor suppressor gene, hereinafter referred to as the "CATR1 gene". The method comprises isolating mRNA from tissue samples, amplifying the mRNA by reverse transcriptase-PCR using primers specific to a region in the CATR1 gene, and detecting the presence or absence of the amplified product to determine whether CATR1 mRNA is present or absent or present at reduced levels in the tissue sample. Optionally, the CATR1 mRNA when present, is also quantified. The present invention also relates to the primers which are used in the method. The present invention also relates to a segment of the CATR1 gene, hereinafter referred to as the "CATR 1.3 genetic element," which is useful for designing the primers used in the method of detecting CATR1 mRNA. The CATR 1.3 genetic element is also useful for preparing antisense nucleic acid segments which are CATR1 gene specific inhibitors.

Abstract: A method for electrophoresis of nucleic acid fragments present in the solution which contains an amount, e.g., 0.2% or more, of a reagent, e.g., glycerol, dithiolthreitol (DTT) and trehalose or other sugars, which interact to form a complex with borate or boric acid. The method includes applying the solution to an electrophoretic gel and electrophoresing those fragments into the gel in the presence of a buffer lacking boric acid, or a derivative thereof, which forms a chelate complex with the reagent and thereby causes distortion of electrophoresis of the fragments in a gel including such a buffer.

Abstract: Isolated mpl ligand, isolated DNA encoding mpl ligand, and recombinant methods of preparing mpl ligand are disclosed. These mpl ligands are shown to influence the replication, differentiation or maturation of blood cells, especially megakaryocyte progenitor cells. Accordingly, these compounds are used for treatment of thrombocytopenia.

Abstract: A method of identification of differentially expressed messenger RNA (mRNA) which consists of synthesizing from a set of sequences of mRNA sets of fragments of complementary DNA (cDNA), which are separated with the aid of gel electrophoresis and the pictures of separation of the cDNA from different types of cells are compared and fragments with the aid of restriction nucleases. A method of cloning of differentially expressed mRNAs consists of synthesizing from sets of sequences of MRNAs from different types of cells sets of fragments of complementary DNA (cDNA) which are separated with the aid of gel electrophoresis, the pictures of the separation of the cDNA from different types of cells are compared, fragments of cDNA with different signal intensities are separated from the gel, amplified with the aid of a polymerase chain reaction and clones to a plasmid or phage vector.

Abstract: The present invention discloses deoxyribonucleic acid enzymes--catalytic or enzymatic DNA molecules--capable of cleaving nucleic acid sequences or molecules, particularly RNA, in a site-specific manner, as well as compositions including same. Methods of making and using the disclosed enzymes and compositions are also disclosed.

Abstract: The invention provides methods for the detection of mutations and polymorphisms in the pRb2/p130 gene, which may be used to characterize genetic events associated with tumor formation, to trace the parental origin of mutatations, to identify carriers of germline mutations, and to identify individuals with a predisposition to cancer.

Abstract: Telomerase activity in a sample can be measured using a two reaction protocol. The first reaction involves the formation of telomerase substrate extension products from a telomerase substrate. The second reaction involves replication of the telomerase substrate extension products and/or amplification of signal generated by a bound probe. The presence of telomerase activity in a human somatic tissue or cell sample is positively correlated with the presence of cancer and can be used to diagnose a disease or other conditions of medical interest, as well as the course of disease progression or remission in a patient.

Abstract: Described are methods and means for the construction and microbial expression of quasi-synthetic genes arising from the combination of organic synthesis and enzymatic reverse transcription from messenger RNA sequences incomplete from the standpoint of the desired protein product. Preferred products of expression lack bio-inactivating leader sequences common in eukaryotic expression products but problematic with regard to microbial cleavage to yield bioactive material. Illustrative is a preferred embodiment in which a gene coding for human growth hormone (useful in, e.g., treatment of hypopituitary dwarfism) is constructed and expressed.