Abstract: The present invention provides a fermentation process for improving the production level of a recombinant human collagen. The process comprises inoculating a sterilized fermentation medium with a Pichia yeast solution, then performing fermentation culturing for 14-18 hrs, adding methanol to induce the expression, upon which sodium pyruvate is added to the fermentation medium, wherein the amount of the sodium pyruvate added is 0.01-10 g/L. In the fermentation process of the present invention, the sodium pyruvate is added in the methanol-induced expression stage, so that the biosynthesis rate of the recombinant human collagen is improved; and a continuous feed-batch mode is adopted, so that the biosynthesis rate of the recombinant human collagen is further improved, and the fermentation time is shortened. Meanwhile, the expression level of the recombinant human collagen is increased, the fermentation level is increased by more than 20%, and the production cost is reduced.

Abstract: The present invention discloses a fermentation process with a Pichia yeast expressing a recombinant protein, in which Pichia pastoris is used as a fungal strain. The process comprises performing primary seed culturing to reach a fungal concentration of 20±2 g/L; then performing secondary seed culturing to reach a fungal concentration of 120±10 g/L; next proceeding to a glycerol culturing stage, wherein the amount of glycerol added in the glycerol culturing stage is 60-70 g/L; and then proceeding to a methanol-induced stage for 120±8 h after the dissolved oxygen quickly reaches a relatively stable state, to complete the fermentation process. In the present invention, a glycerol fed-batch addition stage in existing processes is omitted, and the process proceeds to a next stage as soon as glycerol is completely consumed, with no need to prepare sterilized glycerol for fluidic addition. As such, the glycerin sterilizer is omitted, the consumption of energy and resource and the waste of glycerin are reduced.

Abstract: The present invention discloses a fermentation process with a Pichia yeast expressing a recombinant protein, in which Pichia pastoris is used as a fungal strain. The process comprises performing primary seed culturing to reach a fungal concentration of 20±2 g/L; then performing secondary seed culturing to reach a fungal concentration of 120±10 g/L; next proceeding to a glycerol culturing stage, wherein the amount of glycerol added in the glycerol culturing stage is 60-70 g/L; and then proceeding to a methanol-induced stage for 120±8 h after the dissolved oxygen quickly reaches a relatively stable state, to complete the fermentation process. In the present invention, a glycerol fed-batch addition stage in existing processes is omitted, and the process proceeds to a next stage as soon as glycerol is completely consumed, with no need to prepare sterilized glycerol for fluidic addition. As such, the glycerin sterilizer is omitted, the consumption of energy and resource and the waste of glycerin are reduced.

Abstract: The present invention provides a fermentation process for improving the production level of a recombinant human collagen. The process comprises inoculating a sterilized fermentation medium with a Pichia yeast solution, then performing fermentation culturing for 14-18 hrs, adding methanol to induce the expression, upon which sodium pyruvate is added to the fermentation medium, wherein the amount of the sodium pyruvate added is 0.01-10 g/L. In the fermentation process of the present invention, the sodium pyruvate is added in the methanol-induced expression stage, so that the biosynthesis rate of the recombinant human collagen is improved; and a continuous feed-batch mode is adopted, so that the biosynthesis rate of the recombinant human collagen is further improved, and the fermentation time is shortened. Meanwhile, the expression level of the recombinant human collagen is increased, the fermentation level is increased by more than 20%, and the production cost is reduced.

Abstract: The present invention provides a method of site-directed insertion to H11 locus in pigs by using site-directed cutting system, includes the following steps: 1) identify the targeted sequence targeted by the targeted cutting system in the targeted genome sequence of pigs; 2) design and construct the targeting sequence of the corresponding cutting system according to the targeted site; 3) construction of targeting vector; 4) transfect cells, identify the efficiency of fixed-point insertion by PCR amplification. The invention is dependent on the site-directed cutting system of H11 locus in pigs, to insert the target gene into the target site, in order to solve the problems such as low efficiency of traditional shooting technique, inconvenience design of PCR detection primer, harder to detect. The invention provides a method of site-directed insertion which can stably express the foreign gene at the H11 locus, to build an efficient platform for the production of transgenic pigs.

Abstract: A method for auxiliary identification of WZSP inbred line and its special primers are disclosed. The method may comprise: testing whether a deoxyribose nucleotide at site 1273 from 5? end of genomic DNA comprising SEQ ID NO: 1 in a test pig is A or G, measuring whether the test pig genotype is GG, GA or AA wherein the test pig with GG genotype is a candidate for the WZSP inbred line and the test pigs with GA genotype and AA genotype are non-WZSP inbred line. The method can be applied to breed WZSP inbred line, which can be used to preliminarily screen all the pigs in test pig group, eliminate non-WZSP inbred line, find out a candidate WZSP inbred line and make a further confirmation in combination with other methods. The method and primers can also be used to judge whether a WZSP on market is counterfeiting or not.

Abstract: A method for preparing a transgenic animal of simultaneous multiple-gene expression is provided. Additionally, a method for preparing a transgenic embryo, which introduces both phytase gene and human myxovirus resistant gene A into a target embryo, to obtain a transgenic embryo is provided. The transgenic animal of simultaneous multiple-gene expression can be achieved by transplanting the transgenic embryo into the body of a female target animal. A significant advantage of the foregoing methods, among many others, exists in that the simultaneous expression of multiple genes can be achieved in one transgenosis, which provides a convenient method for the preparation of combined-gene transferred animals.

Abstract: A method, system, and computer-readable storage medium are disclosed for aligning user scribbles to edges in an image. A plurality of edges in the image may be determined. User input comprising a scribble may be received, wherein the scribble comprises a freeform line overlaid on the image. The scribble may be automatically aligned to one or more of the edges in the image.

Abstract: A method, system, and computer-readable storage medium are disclosed for aligning user scribbles to edges in an image. A plurality of edges in the image may be determined. User input comprising a scribble may be received, wherein the scribble comprises a freeform line overlaid on the image. The scribble may be automatically aligned to one or more of the edges in the image.

Abstract: A method for cultivating a transgenic animal with an increased expression amount of porcine growth hormone is provided. Additionally, a method for cultivating a transgenic embryo, which introduces a recombinant expression vector of 1) TRE-GH or 2) TET-ON into a target embryo to obtain a transgenic embryo with a higher expression amount of porcine growth hormone than a target embryo without introduction is provided. A transgenic animal can be obtained by transplanting the embryo into an animal. The experiments demonstrate that growth hormone (GH) is sharply increased in blood of transgenic pigs after adding DOX.

Abstract: An auxiliary method for identification of WZSP inbred line and its special primers are provided. The method is to test if DNA at site 1273 from 5? end of genomic DNA SEQ ID NO: 1 in test pig is A or G, measure the test pig genotype is GG, GA or AA; GG or AA genotype is homozygote that DNA at site 1273 is G or A; GA genotype is their heterozygote; test pig of GG genotype is candidate for the WZSP inbred line; test pigs of GA and AA genotypes are not WZSP inbred line. The method can be applied to breed WZSP inbred line, can preliminarily screen all the pigs in test pig group, eliminate non WZSP inbred line, find out candidate WZSP inbred line, combine with other methods for further confirmation. The method can also be used to judge whether WZSP on market is counterfeiting or not.

Abstract: A method for preparing a transgenic animal of simultaneous multiple-gene expression is provided. Additionally, a method for preparing a transgenic embryo, which introduces both phytase gene and human myxovirus resistant gene A into a target embryo, to obtain a transgenic embryo is provided. The transgenic animal of simultaneous multiple-gene expression can be achieved by transplanting the transgenic embryo into the body of a female target animal. A significant advantage of the foregoing methods, among many others, exists in that the simultaneous expression of multiple genes can be achieved in one transgenosis, which provides a convenient mean for the preparation of combined-gene transferred animals etc.

Abstract: A frame container for shipping chassis includes a base frame and corner posts. The base frame includes two bottom side beams each extending in a longitudinal direction between two corner posts and beams extending between two bottom side beams in a direction that is perpendicular to the bottom side beams. Each bottom side beam includes an upper surface and a lower surface extending in the longitudinal direction between the two corner posts and a locking hole defined between the upper and lower surfaces for receiving a twist lock of a chassis.

Abstract: A method used for loading semi-trailer chassis into a container, wherein the container includes a base frame and four frame structures attached to corner posts of the base frame. Four semi-trailer chassis are stacked into the container from bottom to top, where the first semi-trailer chassis is placed upside down, the second semi-trailer chassis is placed right-side up and turned horizontally at a 180 degree angle, the third semi-trailer chassis is placed upside down, and the fourth semi-trailer chassis is placed right-side up and turned horizontally at a 180 degree angle.

Abstract: The present invention provides an aerator of a container comprising an aerator mounted on the side board of the container and an electronic label installed in the interior space of the aerator. The present invention also provides a container comprising a container body, a penetrated hole locating on the side board of the container body, and an aerator installed in the penetrated hole from the outside of the container body. An electronic label is installed in the interior space of the aerator. The electronic label based on the RFID technology is combined with the aerator of the container. Since the electronic label stores the relative data of the containers which can be read by the readers through radio, the container will be automatically identified.

Abstract: A method used for loading of container shipping semi-trailer chassis, in which container mentioned includes base frame and four frame structures fixed on angle post of base frame; four semi-trailer chassis are loaded into the container in the order from bottom to top according to positioning of the first semi-trailer chassis being placed upside down, the second one being placed upside up and turned by 180 degree horizontally, the third one being placed upside down, the forth one being placed upside up and turned by 180 degree horizontally.

Abstract: A container used for transportation of semi-trailer chassis, frame structure is adopted for the container mentioned, including: at least two pieces of bottom side beam; at least two pieces of bottom cross beam intersecting with bottom side beam; at least four pieces of vertical angle post intersecting with bottom side beam or bottom cross beam; bottom side beam, bottom cross beam and angle post are connected to form a rectangle frame structure. The container is specially used to ship semi-trailer chassis, it can load four stacked semi-trailer chassis, therefore realize containerized transportation of semi-trailer chassis, reaching the target of enhancing of efficiency of transportation of semi-trailer chassis and reduction of transportation cost of semi-trailer chassis.