Specific primers and typing method of class II MHC (major histocompatibility complex) genes for antibacterial potential detection of alligator sinensis

A typing method and technology of Chinese alligator, applied in DNA/RNA fragments, recombinant DNA technology, etc.

Inactive Publication Date: 2016-02-10
ZHEJIANG UNIV
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Problems solved by technology

[0005] In the existing Chinese alligator MHC literature, the cross-amplification of the MHC gene sites is performed using universal primers, and the results can only be used for simple evolutionary analysis

Method used

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  • Specific primers and typing method of class II MHC (major histocompatibility complex) genes for antibacterial potential detection of alligator sinensis
  • Specific primers and typing method of class II MHC (major histocompatibility complex) genes for antibacterial potential detection of alligator sinensis
  • Specific primers and typing method of class II MHC (major histocompatibility complex) genes for antibacterial potential detection of alligator sinensis

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Embodiment Construction

[0027] A pair of primers of the present invention for amplifying the gene sequence of the alligator class II major histocompatibility complex (MHC) is based on the MHC gene fragment obtained by BAC sequencing in the previous stage, according to the intron of the MHC gene For the characteristics of large differences, use PrimerPremier5 to design site-specific primers. Many pairs of primers have been designed in the process, and the experimental results show that the primers of the present invention are the best.

[0028] Apply primers of the present invention to amplify Chinese alligator MHC gene and analyze the specific method as follows:

[0029] 1. Extract the DNA of the Chinese alligator blood sample to be tested by phenol-chloroform extraction method as a template;

[0030] 2. Perform PCR amplification on the extracted DNA. The 10 μL PCR reaction system is: 0.8 μL of Chinese alligator blood genomic DNA, 0.2 μL of upstream and downstream primers, 4.4 μL of ultrapure water,...

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Abstract

The invention relates to a pair of single-locus specific primers for amplifying class II major histocompatibility complex genes for antibacterial potential detection of the alligator sinensis and a typing method, wherein the specific sequences of the primers are as shown in SEQ ID NO. 1-SEQ ID NO. 2. The amplification primers disclosed by the invention are stable in single-locus specific amplification capacity in an alligator sinensis population; a product obtained from amplification can conduct genotyping on various individuals through a subsequent single strand conformation polymorphism (SSCP) experiment, so that the polymorphism of the class II MHC genes of alligator sinensis individuals in the population is assessed and the potential of the alligator sinensis individuals in resisting bacterial diseases is detected; therefore, an important reference is provided for such operations of new population founder selection, reintroduction individual selection, artificial mating selection and the like in alligator sinensis protection work.

Description

technical field [0001] The invention discloses a pair of single-site specific primers and a typing method for amplifying a class II major histocompatibility complex (MHC) gene sequence for detecting the antibacterial potential of Chinese alligator. Background technique [0002] Chinese alligator (Alligators inensis, Fauvel) belongs to the class Reptilia, Crocodiles, Fauvelidae, Alligator subfamily, Fauvel genus, is an ancient animal, originated in the Triassic period 230 million years ago, known as reptiles The "living fossil" in animals is a rare and endangered animal unique to my country and a national first-level key protected animal (1972). It was listed in Appendix I of the Convention on International Trade in Endangered Animals and Plants (CITES), and was also recognized by the International Union for Conservation of Nature ( IUCN) is classified as a critically endangered species and is one of the most threatened and endangered species of crocodilians in existence. At p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858
Inventor 方盛国万秋红翟腾
Owner ZHEJIANG UNIV
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