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Method of increasing yield of human papilloma virus L1 albumen pronucleus expression

A technology for human papillomavirus and L1 protein, which is applied in the field of prevention and treatment of human papillomavirus infection, can solve problems such as reduced expression efficiency, and achieve the effects of increasing expression amount, reducing costs, major economic benefits and social benefits

Active Publication Date: 2007-08-15
BEIJING HEALTH GUARD BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when Escherichia coli expresses recombinant genes of higher mammals and humans, the expression efficiency is sometimes reduced due to the differences in the microenvironment of protein translation modification between eukaryotic and prokaryotic expression

Method used

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  • Method of increasing yield of human papilloma virus L1 albumen pronucleus expression
  • Method of increasing yield of human papilloma virus L1 albumen pronucleus expression
  • Method of increasing yield of human papilloma virus L1 albumen pronucleus expression

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Example 1 carries out codon optimization to the nucleic acid sequence encoding HPV16 L1 protein

[0046] In the examples of the present invention, all primer synthesis and DNA sequence determination were completed by Huamei Biotechnology Co., Ltd. For routine operations such as base replacement, SDS-polyacrylamide gel electrophoresis, and subcloning, please refer to "Molecular Cloning" (p. Third edition, Science Press, published in August 2002).

[0047] The specific method is as follows:

[0048] 1. According to the wild HPV16 L1 gene sequence (Genebank accession number AF402678), design a set of oligonucleotide primers that overlap each other end to end, and the overlapping parts are paired according to the principle of double-stranded DNA base complementarity, thus serving as templates for each other, until all are covered Reading frame of HPV16L1 DNA sequence. The average base length of each primer is 60 bases.

[0049] 2. For the DNA primer sequence of each HPV1...

Embodiment 2

[0057] Example 2 Carry out codon optimization to the nucleic acid sequence encoding L1 protein of the remaining HPV types

[0058] Operation steps and detection of codon-optimized nucleic acid sequences encoding L1 protein of other HPV types (including HPV6, HPV11, HPV18, HPV31, HPV33, HPV35, HPV39, HPV45, HPV51, HPV52, HPV 56, HPV58, HPV59, HPV68) The method is all the same as in Example 1, except that in step 1, according to the wild gene sequences of different HPV types (the corresponding Genebank accession number sees the technical scheme part), the oligonucleotides designed to overlap each other end to end, and the overlapping parts are templates for each other Nucleotide primers are different, so that the remaining operations will be changed accordingly according to the different oligonucleotide primers. These changes are all well-known techniques for those skilled in the art. The codon optimized sequences obtained are shown in Figure 1- 15.

[0059] As detected by SDS-...

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Abstract

The invention discloses a method to increase human papilloma virus L1 protein pronucleus expression productivity and also discloses a encode HPV L1 protein codon majorizing nucleic acid sequence from this method, which is characterized by the following: supplying expression carrier and host cell and HPVL1 protein poly body of nucleic acid sequence; disclosing appliance in preparing vaccine, drug compound and immunodiagnosis or antibody. The nucleic acid sequence expressing quantity possesses distinctive improvement, which decreases the preparing cost effectively.

Description

technical field [0001] The present invention relates to the field of prevention and treatment of human papillomavirus infection. Specifically, the present invention relates to a method for improving the prokaryotic expression yield of human papillomavirus L1 protein, and the codon-optimized nucleic acid sequence encoding human papillomavirus L1 protein obtained according to the method, the vector and host comprising these nucleic acid sequences cells, and the present invention also relates to the HPV L1 protein multimer expressed in Escherichia coli by the codon-optimized nucleic acid sequence, and the application of the HPV L1 protein multimer in the preparation of vaccines, pharmaceutical compositions, diagnostic antigens or antibodies. Background technique [0002] In the field of bioengineering and biopharmaceuticals, using Escherichia coli as a host cell to express recombinant protein products is more economical and effective than using eukaryotic host cells such as yea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12N15/70C07K14/025A61K39/12A61P31/12G01N33/53G01N33/68G01N33/569
Inventor 马润林陈小江
Owner BEIJING HEALTH GUARD BIOTECH
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