Abstract: In accordance with the present invention, there is provided a novel restriction endonuclease and its DNA obtainable from Helicobacter pylori J188 (NEB#1174), hereinafter referred to as “Hpy188I”, which endonuclease: (1) recognizes the nucleotide sequence 5′-TCNGA-3′ in a double-stranded DNA molecule as shown below, 5′-TCN↓GA-3′ 3′-AG↑NCT-5′ (wherein G represents guanine, C represents cytosine, A represents adenine, T represents thymine and N represents either G, C, A, or T); (2) cleaves said sequence in the phosphodiester bonds between the N and G as indicated with the arrows; and (3) cleaves double-stranded PhiX174 DNA to produce 18 fragments, including fragments of 1331, 813, 572, 525, 396, 302, 293, 219 base pairs, and 10 fragments smaller than 200 base pairs.

Abstract: The present invention relates to the recombinant DNA which encodes the SwaI restriction endonuclease, modification methylase, and the production of SwaI restriction endonuclease from the recombinant DNA. Related expression vectors, pHKUV5 which features a strong, constitutive UV5 promoter without the Lac repressor binding site and pHKT7 which contains a powerful controllable T7 promoter and a low copy number origin of replication, are also disclosed.

Abstract: In accordance with the present invention, there is provided a novel restriction endonuclease and its DNA obtainable from Helicobacter pylori CH4 (NEB#1236), hereinafter referred to as “HpyCH4III”, which endonuclease: (1) recognizes the nucleotide sequence 5′-ACNGT-3′ in a double-stranded DNA molecule as shown below, 5′-ACN↓GT-3′ 3′-TG↑NCA-5′  (wherein G represents guanine, C represents cytosine, A represents adenine, T represents thymine and N represents either G, C, A, or T); (2) cleaves said sequence in the phosphodiester bonds between the N and G as indicated with the arrows; and (3) cleaves double-stranded PhiX174 DNA to produce 15 fragments, including fragments of 1284, 814, 536, 517, 454, 404, 302, 292, 270 and 222 base pairs, and 5 fragments smaller than 200 base pairs.

Abstract: In accordance with the present invention, there is provided a novel restriction endonuclease and its DNA obtainable from Helicobacter pylori J188 (NEB#1174), hereinafter referred to as “Hpy188III”, which endonuclease: (1) recognizes the nucleotide sequence 5′-TCNNGA-3′ in a double-stranded DNA molecule as shown below, 5′-TC↓NNGA-3′ 3′-AGNN↑CT-5′ (wherein G represents guanine, C represents cytosine, A represents adenine, T represents thymine and N represents either G, C, A, or T); (2) cleaves said sequence in the phosphodiester bonds between the C and first unspecified nucleotide N as indicated with the arrows; and (3) cleaves double-stranded PhiX174 DNA to produce 26 fragments, including fragments of 704, 560, 497, 477, 423, 309, 308, 304, 261, 225 basepairs, and 16 fragments smaller than 200 basepairs.

Abstract: RsaI, a restriction enzyme from the bacterium Rhodopseudomonas sphaeroides, recognizes the DNA sequence 5′-GTAC-3′. Because RsaI is commercially valuable, we sought to overproduce it by cloning the genes for RsaI and its accompanying, modification, enzyme. The ‘methylase-selection’ method, the customary procedure for cloning restriction and modification genes, was applied to RsaI. The method yielded clones containing the methylase gene (rsaIM), but none containing both the methylase gene and the restriction gene (rsaIR). Inverse-PCR was then used to recover sections of the DNA downstream of rsaIM. These sections were sequenced, and the sequences were joined in silico to reveal the gene organization of the RsaI R-M system.

Abstract: The present invention relates to cloned DNA containing origin of DNA replication and to cloned DNA encoding repliation protein, RepT.

Abstract: In accordance with the present invention, there is provided a novel restriction endonuclease and its DNA obtainable from Helicobacter pylori CH4 (NEB#1236), hereinafter referred to as “HpyCH4IV”, which endonuclease: (1) recognizes the nucleotide sequence 5′-ACGT-3′ to in a double-stranded DNA molecule as shown below, 5′-A↓CGT-3′ 3′-TGC↑A-5′ (wherein G represents guanine, C represents cytosine, A represents adenine, T represents thymine and N represents either G, C, A, or T); (2) cleaves said sequence in the phosphodiester bonds between the A and C as indicated with the arrows; and (3) cleaves double-stranded PhiX174 DNA to produce 19 fragments, including fragments of 1036, 749, 397, 379, 371, 365, 362, 343, 282, 282, 268 base pairs, and 8 fragments smaller than 200 base pairs.

Abstract: The present invention relates to recombinant DNA which encodes a novel nicking endonuclease, N.BstNBI, and the production of N.BstNBI restriction endonuclease from the recombinant DNA utilizing PleI modification methylase. Related expression vectors, as well as the application of N.BstNBI in non-thio strand displacement amplification, is disclosed also.

Abstract: The present invention relates to the use of these cyclophilins, hereinafter referred to as `tyrosine-containing` cyclophilins, in a method for identifying compounds capable of binding to and/or inhibiting the enzymatic activity of these proteins. Such compounds may be further screened for their ability to inhibit parasites which are not susceptible to the anti-parasitic effects of CsA.

Abstract: A genomic DNA library of Thermus filiformis was constructed using pBR322 as a cloning vector. The methylase selection method was used to clone the TfiI methylase gene (tfiIM). A clone carrying an active TfiI methylase was identified. After sequencing the complete TfiI methylase gene and its downstream DNA sequence, a recombinase homolog was found. Because the methylase and its cognate endonuclease gene are located in proximity to each other in a particular restriction-modification system, efforts were made to clone the upstream DNA by inverse PCR. After two rounds of inverse PCR, one open reading frame (ORF1) was found upstream of the TfiI methylase gene. This ORF1, containing a Shine-Dalgarno sequence and a TATA box on the upstream side, was cloned and expressed, and TfiI endonuclease activity was detected in crude cell extracts. It is concluded that ORF1 encodes TfiI restriction endonuclease.

Abstract: The present invention relates to recombinant DNA molecules encoding NspHI restriction endonuclease and methylase and to method to use premodified E. coli K strain RR1 (.lambda.DE3) for overexpression of NspHI restriction endonuclease.

Abstract: The present invention relates to the use of nematode "cyclophilin-like proteins (CLP)", in a method for identifying compounds capable of binding to and/or inhibiting the peptidyl-prolylcistransisomerase activity of these proteins. Such compounds may be further screened for their ability to inhibit nematode parasites which are not susceptible to the anti-parasitic effects of CsA.

Abstract: In accordance with the present invention, a 31-33 kDa glycoprotein of D. immitis (DiT33) is provided which represents another member of the family of putative pepsin inhibitors. Other known members include Ov33 (a.k.a. Ov33.3, Oc3.6, OvD 5B), Bm33 and Av33. These filarial molecules possess significant homology to the known pepsin inhibitor (Aspi3) of A. suum. Using DiT33 or Ov33, in the form of a recombinant fusion or non-fusion protein, antibody responses to DiT33 may be monitored and used in immunodiagnosis of heartworm infection in mammals. Antibodies reactive with the DiT33 or Ov33 may also be used to detect DiT33 antigen as a means of immunodiagnosis of heartworm infection in mammals.

Abstract: BsrFI restriction enzyme was purified from Bacillus stearothermophilus to near homogeneity. The protein was sequenced to obtain its N-terminus amino acid sequence. A set of denegerate primers were synthesized based on the aa sequence. The first 18 codons encoding BsrFI restriction endonuclease (bsrFIR) was amplified by PCR and its coding sequence was obtained. The methylase selection method was used to clone BsrFI methylase gene (bsrFIM). Two clones were found to be resistant to BsrFI digstion. The entire insert in one clone was sequenced and the insert encodes the BsrFI methylase (M. BsrFI). In addition, a small truncated open reading frame adjacent to the methylase gene has homology to Cfr10I restriciton endonuclease in a BlastX homology search in Genbank database. BsrFI and Cfr10I are isoschizomer that recognizes and cleaves 5'R CCGGY3'.

Abstract: The present invention relates to the recombinant DNA which encodes the SgrAI restriction endonuclease and the MspI modification methylase, and the production of SgrAI restriction endonuclease from the recombinant DNA.

Abstract: The present invention relates to the recombinant DNA which encodes the DraIII restriction endonuclease modification methylase, and the production of DraIII restriction endonuclease from the recombinant DNA. Related expression vectors, pHKUV5 vector which features a strong, constitutive UV5 promoter without the Lac repressor binding site and pHKT7 vector which contains a powerful controllable T7 promoter and a low copy number origin of replication, are also disclosed.

Abstract: The present invention relates to clones which express the recombinant NspI restriction endonuclease using a NspI methylase premodified E. coli K strain RR1 (.lambda.DE3) and to methods for producing and purifying said enzyme.

Abstract: The methylase selection method was used to clone the SnaBI methylase gene (snaBIM) from Sphaerotilus natans (ATCC 15291). An active SnaBI methylase was cloned in E. coli using pSnaBI-2, a pUC19 derivative containing two SnaBI sites. Because methylase and restriction genes are usually located alongside each other in a restriction-modification systems, efforts were made to clone the downstream DNA by inverse PCR. Inverse PCR cloned the missing portion of the SnaBI endonuclease and also identified a control, or C, protein. The two open reading frames snaBIR (ORF1) and snaBIC (ORF2) converged toward the SnaBI methylase gene (ORF). Attempts to establish a snaBIR-recombinant plasmid expressing the SnaBI endonuclease in E.coli modified with SnaBI methylase failed. Overexpression of the SnaBI endonuclease in E. coli required the use of the heterospecific BsaAI methylase.

Abstract: A recombinant restriction endonuclease from Anabaena variabilis is provided, as well as the isolated gene which encodes it and methods for the production of the recombinant AvaI restriction endonuclease. An isolated gene encoding a modification methylase from A. variabilis is also provided.

Abstract: The present invention relates to isolating DNA coding for DNA polymerase I from Thermomicrobium roseum, expressing the T. roseum DNA polymerase I gene in E. coli and purifying the recombinant T. roseum DNA polymerase I from E. coli cell extract.