Method for constructing recombinant plasmid of human papilloma virus subtype L1 gene

Abstract

The invention discloses a method for constructing recombinant plasmid of a human papilloma virus (HPV) subtype L1 gene. The problem that the HPV detection result of interference performance is liableto occur due to lack of various subtype L1 positive control nucleic acid standard samples at present is solved. The method for constructing the recombinant plasmid of the human papilloma virus (HPV) subtype L1 gene is characterized in that an NCBI database finds a gene sequence of different subtype L1 areas of HPV; the gene sequence is subjected to bioinformatics analysis by a BLAST function on awebsite; a GP5<+>/6<+> area is selected as a target gene target fragment; 25 groups of specific primers are designed separately by using Primer Premier 5.0 software; a corresponding positive sample issubjected to PCR amplification; a amplification product is used for establishing standard plasmid, construction work of 25 subtype recombinant standard plasmid of the HPV is finished successfully; and guarantee is provided for the stability of the test result with high-risk HPV infection or not which is screened by a molecular biological technology of HPV nucleic acid detection.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for constructing a recombinant plasmid of human papillomavirus subtype L1 gene. Background technique [0002] Human papillomavirus (HPV), belonging to the Papillomavirus genus of the family Papovaviridae, is a small-molecule, non-enveloped virus. The HPV genome is a circular double-stranded DNA with a length of about 8000 base pairs (bp). [0003] According to the function, the HPV genome can be divided into three functional regions: (1) Early transcription region: also known as the E region, consisting of 4500 base pairs, encoded as E1, E2, E3, E4, E5, E6, E7 and E8 are eight early proteins, among which the function of E1 is closely related to the replication of HPV DNA; E2 is a type of activator protein, E5 is closely related to cell transformation, and the early E6 and E7 are the main cancers on the surface of HPV Gene, so the early router mainly has the...

AI Technical Summary

Problems solved by technology

[0008] Most of the existing molecular biology techniques for HPV nucleic acid detection are aimed at the L1 conserved region in the genome of the HPV virus, but at present, due to the lack of L1 positive control nucleic acid standard samples of various subtypes, interference-like HPV detection results (false negatives) are prone to occur. or false positive), false negative results may lead to untimely diagnosis and treatment of cervical cancer, false positive results may lead to unnecessary frequent screening and invasive treatment, which will cause potential public health risks

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