KR910012243A - Human tumor necrosis factor (TNF-α) expression vector [pYLBC-A / G-TNF-α] and preparation method thereof - Google Patents

Abstract

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Description

Human tumor necrosis factor (TNF-α) expression vector [pYLBC-A / G-UB-HGH] and its preparation method

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3 shows the cloning process into a yeast expression carrier according to the present invention.

Claims (8)

After confirming the nucleotide sequence of human tumor necrosis factor in the cDNA library, a human tumor necrosis factor gene having restriction enzymes Bgl II and Xba I Sal I recognition sites was prepared, and then the gene was conjugated with an amino terminal synthesis gene, and the effector gene and the termination gene. Cloning the E. coli carrier containing the yeast agonist gene, human tumor necrosis factor gene and the end codon to obtain a cassette, and then cloned into the yeast expression vector pYLBC human tumor necrosis factor expression vector, characterized in that pYLBC-A / G-TNF-α]. The human tumor necrosis factor gene of claim 1 has restriction enzyme Bgl II sites at the 3rd and 4th amino acid positions of human tumor necrosis, and has restriction enzyme Xba I sites at the 81st and 82th amino acid positions, and the carboxy terminal. Human tumor necrosis factor expression vector [pYLBC-A / G-TNF-α], characterized in that it has a restriction enzyme Sal I site at the position. [4] The human tumor necrosis factor expression vector [pYLBC-A] according to claim 1, wherein the amino-terminal synthetic gene is complementary to the restriction enzyme Nco I site at the start codon, and the opposite side to the restriction enzyme Bgl II site. / G-TNF-α]. 4. The amino terminus synthetic gene of claim 1 or 3, wherein the amino terminus synthetic gene is comprised of an upper strand 8-mer (5'-CATG GTTA-3 ') and a lower strand 8-mer (3'-CAA TCT AG-5'). Human tumor necrosis factor expression vector [pYLBC-A / G-TNF-α]. After confirming the nucleotide sequence of human tumor necrosis factor in the cDNA library, a human tumor necrosis factor gene having restriction enzymes Bgl II and Xba I Sal I recognition sites was prepared, and then the gene was conjugated with an amino terminal synthesis gene, and the effector gene and the termination gene. Cloning the E. coli carrier having a yeast working gene, a human tumor necrosis factor gene and a cassette consisting of the end codon, and then recloning it into the yeast expression carrier human tumor necrosis factor expression vector [pYLBC- A / G-TNF-α]. The human tumor necrosis factor gene of claim 5 has a restriction enzyme Bgl II site at the 3rd and 4th amino acid positions of human tumor necrosis and has a restriction enzyme Xba I site at the 81st and 82nd amino acid positions, and A method for producing a human tumor necrosis factor expression vector [pYLBC-A / G-TNF-α], which has a restriction enzyme Sal I site at a carboxy terminal position. [6] The human tumor necrosis factor expression vector [pYLBC-A] according to claim 5, wherein the amino-terminal synthetic gene is complementary to the restriction enzyme Nco I site at the start codon, and the ATC site is complementary to the restriction enzyme Bgl II site. / G-TNF-α]. The method of claim 5 or 7, wherein the amino terminal synthesis gene is composed of the upper 8-mer (5'-CATG GTTA-3 ') and the lower strand 8-mer (3'-CAA TCT AG-5'). A method for producing a human tumor necrosis factor expression vector [pYLBC-A / G-TNF-α]. ※ Note: The disclosure is based on the initial application.