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Deletion mutant of a set of hepatitis B HBx gene and application thereof

A deletion mutant, hepatitis B technology, applied in application, gene therapy, genetic engineering, etc., can solve the problems of clinical diagnosis and treatment without HBx gene deletion mutant

Inactive Publication Date: 2009-10-28
朱平安
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

So far, there have been no reports of HBx gene deletion mutations directly causing HCC, and no reports of HBx gene deletion mutants being used for clinical diagnosis and treatment.

Method used

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  • Deletion mutant of a set of hepatitis B HBx gene and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0013] Example 1: Deletion Mutant Sequence Determination and Application Research

[0014] 1. Extraction of total DNA from liver cancer tissue

[0015] 63 liver cancer tissue samples were obtained from outpatients and inpatients of General Surgery Department and Department of Infectious Diseases of Xiangya Hospital of Central South University. The patients were from Hunan, Hubei, Jiangxi and other provinces. The patients were positive for HBsAg, but negative for anti-HBs, anti-HCV, and anti-HDV triple antibodies . After surgical operation of these liver cancer tissues, the DNA extraction kit of Beijing Tiangen Biotechnology Co., Ltd. was used to extract the DNA of the liver cancer tissues.

[0016] 2. Nested PCR, single-strand conformation polymorphism analysis (SSCP) and heteroduplex analysis (HA)

[0017] 1) Nested PCR: Nested PCR was used to amplify the HBx gene fragment in the liver cancer tissue, and the primer sequences were:

[0018] P1: 5'-ATCGTTTCCATGGCTGCTAGGCT-3'...

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Abstract

The invention provides a group of deletion mutants of hepatitis B HBx gene, wherein SEQ ID NO.1 is a mutant in which 264 bases are deleted between HBV nucleotide positions 1577-1840 in HBV gene; SEQ ID NO.2 is HBx A mutant in which 19 bases are deleted between nucleotide positions 382-400 in the gene; SEQ ID NO.3 is a mutant in which 15 bases are deleted in nucleotide positions 431-445 in the HBx gene. This group of deletion mutants all have obvious tumorigenic effects, and can be used for gene detection of canceration in hepatitis B virus carriers and chronic hepatitis B patients, as well as preparation of drugs for preventing liver cancer and treating HBV chronic infection.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to a group of deletion mutants of hepatitis B HBx gene and its application in clinical diagnosis, treatment and prevention. Background technique [0002] Primary hepatocellular carcinoma (HCC) is one of the most common malignant tumors in Southeast Asia and China. A large number of epidemiological surveys have shown that long-term chronic hepatitis B virus infection is the main cause of HCC. The risk of HCC in people infected with HBV It is more than 100 times that of uninfected patients, and HCC patients have a tendency to cluster HBV infection in their families, suggesting that HBV infection is closely related to the occurrence of HCC. [0003] HBV is a partially double-stranded DNA virus with a genome of 3.2kb, including S, C, P and X4 open reading frames, among which the X gene is the smallest open reading frame and the region with the most obvious functional overlap. The ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/51C12Q1/68A61K48/00A61P1/16A61P35/00
Inventor 朱平安谭德明陈莉彭忠田宋琳刘菲
Owner 朱平安
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