KR880001812A - The structural gene is directly linked to the lower part of the start codon of λPL. Method for preparing E. coli expression vector - Google Patents

Abstract

No content

Description

The structural gene is directly linked to the lower part of the start codon of λPL. Method for producing a collagen expression vector.

As this is a public information case, the full text was not included.

1 is the sequence of the surrounding gene of λP _L

3 is a process for preparing plasmid pSD 100.

5 shows the preparation of plasmid pSD 102 encoding rIFN-αA.

Claims (9)

In the sequence of λP _L , the λP _L transduction plasmid in which the sequence (I) around the start codon was replaced with the sequence (II), (III) or (IV), respectively, was identified as restriction factor KpnI. ΛP by cleavage with Apa I or BstEII, enzymatic digestion of the cleavage sequence at the cleavage site, and insertion of the structural gene at the smooth end, or, optionally, a structural gene having a viscous end complementary to the cleavage sequence at the cleavage site. A method of producing an expression vector of E. coli directly linked to a structural gene directly below the start codon of _L. In this sequence, N is nucleotide A.G.T or C, N 'is a nucleotide complementary to N. The method according to claim 1, wherein when the sequence (I) around the start codon in the sequence of λP _L is replaced with sequence (II), (III) or (IV), sequence (IIa), (IIIa) or (IVa) is used. Way. In said sequence. N and N 'are as defined in claim 1. The method of claim 1, wherein the original sequence (I) is replaced with sequence (II) and the restriction enzyme KpnI is used. The method of claim 1, wherein the original sequence (I) is replaced with sequence (III) and the restriction enzyme ApaI is used. The method of claim 1, wherein the original sequence (I) is replaced with sequence (IV) and the restriction enzyme BstEII is used. The method of claim 1, which is performed using other restriction enzymes that recognize the same site as KpnI, ApaI or BstEII. The method according to claim 1, wherein the sequence containing the sequence (II), (III) or (IV) is replaced by a sequence surrounding the start codon of λP _{L with} a sequence extended to facilitate binding with a structural gene. The method of claim 2, wherein the 3 'end of the coding strand of sequence (IIa), (IIIa) or (IVa) lacks one or more nucleotides than the complementary strand. A method for producing a polypeptide consisting of amino acids that can be genetically encoded by transforming E. coli cells into a vector prepared according to claim 1 and culturing the same, thereby inducing the expression of a gene of interest. ※ Note: The disclosure is based on the initial application.