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PCR method for detecting TRIMCyp genotypes of animals of macaca

A technology for rhesus monkeys and animals, applied in biochemical equipment and methods, microbiological measurement/inspection, etc., can solve unseen problems, achieve the effects of reducing irritation, improving efficiency, and avoiding damage

Inactive Publication Date: 2010-09-01
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI +1
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Problems solved by technology

There are no reports of cynomolgus monkeys carrying the TRIMCyp gene in China

Method used

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  • PCR method for detecting TRIMCyp genotypes of animals of macaca
  • PCR method for detecting TRIMCyp genotypes of animals of macaca
  • PCR method for detecting TRIMCyp genotypes of animals of macaca

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Embodiment

[0021] 1 Materials and methods

[0022] 1.1 Experimental materials The hairs of flat-topped monkeys and cynomolgus monkeys used in the experiment were obtained from the South China Primate Research and Development Center, and the hairs of rhesus monkeys were provided by the Lentivirus Laboratory of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (Note: flat-topped monkeys, cynomolgus monkeys and Rhesus macaque hair can be purchased from macaque farms in various places, for example, from Guangdong Landao Biotechnology Company and Sichuan Yibin Biotechnology Company), and all samples are stored at -20°C until use. Hair Genome Extraction Kit DNA IQ TM System was purchased from Promega. LA Taq enzyme, DNAMarker, and dNTP were all purchased from Bao Biological Engineering (Dalian) Co., Ltd.

[0023] 1.2 Design of primers The TRIM5 gene of macaques contains 8 exons, and CypA in the TRIMCyp genotype of the genus Rhesus macaques has been found to be i...

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Abstract

The invention discloses a PCR method for detecting the mutation of TRIM4 loca of animals of macaca, which comprises the following steps of: (1) extracting DNA from the hair of the animals of the macaca; (2) performing PCR amplification by taking SEQ ID NO:1 and SEQ ID NO:2 as primers; and (3) determining the TRIM5 loca of the animals to be detected are not mutated if an amplification product is one 2,300 to 2,400 bp segment, determining the TRIM5 loca are subjected to heterozygous mutation if the amplification products are the 2,300 to 2,400 bp segment and a 3,000 to 3,150 bp segment respectively, and determining the TRIM5 loca are subjected to homozygous mutation if the amplification product is only the 3,000 to 3,150 bp segment. The method of the invention has the advantage of accurately detecting the TRIMCyp genotypes of the animals of the macaca, and can be used for surveying the distribution of TRIMCyp in macaques in China and screening positive individuals.

Description

technical field [0001] The invention relates to a method for detecting animal genotypes, in particular to a PCR method for detecting TRIMCyp genotypes of rhesus monkeys, belonging to the detection field of TRIMCyp genotypes of rhesus monkeys. Background technique [0002] Current studies have shown that one of the main reasons why HIV-1 cannot replicate efficiently in monkeys is that there are two cellular restriction factors that inhibit retroviruses in monkeys, namely Tripartite motif protein 5-α (TRIM5α) and Apolipoprotein B mRNA-editing catalytic polypeptide 3G (APOBEC3G) (Sakuma R, Noser J A, Ohmine S, Ikeda Y. Inhibition of HIV-1 replication by simian restriction factors, TRIM5[alpha] and APOBEC3G[J]. Gene Ther 2006; 14(2): 185-189 ). This study is limited to TRIM5α, a constitutively expressed cytoplasmic protein that mediates the restriction of retroviral entry into cells. TRIM5α binds to the capsid protein (capsid, CA) of the virus, mainly restricts the shedding of...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 周建华刘晓明于长青饶军华那雷汤艳东吕晓玲胡哲
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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