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Gene full-length sequence of monochamus alternatus triosephosphate isomerase and clone method thereof

A technology of monochamus monochamus and triose phosphate, applied in the direction of isomerase, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2016-03-02
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on Monochamus alternatus mainly focuses on the morphology and structure, comprehensive control, gene expression profile and functional analysis, etc. The research on Monochamus alternatus triose phosphate isomerase and its application in immunology has not been reported yet.

Method used

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  • Gene full-length sequence of monochamus alternatus triosephosphate isomerase and clone method thereof
  • Gene full-length sequence of monochamus alternatus triosephosphate isomerase and clone method thereof
  • Gene full-length sequence of monochamus alternatus triosephosphate isomerase and clone method thereof

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Embodiment Construction

[0024] The following examples further illustrate the invention.

[0025] 1. Extraction of total RNA from Monochamus alternatus

[0026] The adults of Monochamus monochamus were collected from the Pinus massoniana forest in Conghua, Guangzhou. Store in a centrifuge tube after freezing with liquid nitrogen, and store in a -70°C refrigerator for later use.

[0027] Using E.Z.N.A. TM TotalRNAKitII total RNA extraction kit to extract total RNA, the specific method is as follows:

[0028] 1) Grind and pulverize the adult Monochamus alternatus, weigh 100 mg of the sample and grind it into powder in a mortar pre-cooled with liquid nitrogen, and then put it into a 1.5 mL centrifuge tube. Add 1mL RNA-SolvReagent to the centrifuge tube and let stand at room temperature for 3min.

[0029] 2) Add 200 μL of chloroform, shake vigorously for 15 seconds, and incubate on ice for 10 minutes.

[0030] 3) Centrifuge at 12000r / min for 15min at 4°C, and the upper aqueous phase appears.

[0031...

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Abstract

The invention relates to the field of biological gene engineering, in particular to a gene full-length sequence of monochamus alternatus triosephosphate isomerase and a clone method thereof. The clone method includes the steps that monochamus alternatus adults are used for extracting total RNA, RACE amplification is adopted to obtain a gene 3' end sequence, and the gene 3' end sequence and a constructed 5' end sequence of monochamus alternatus triosephosphate isomerase in a cDNA library are spliced to obtain the gene full-length sequence. The full length of the gene sequence is 1450 bp, the length of a gene open reading frame is 514-1257 bp, and 247 pieces of triosephosphate isomerase amino acid are encoded in total. Genes of monochamus alternatus glycolysis enzymes are supplemented, a research thought and a direction are provided for research and development of a novel schistosomiasis vaccine, and the gene full-length sequence is of great significance in immunology.

Description

technical field [0001] The invention relates to the field of biogenetic engineering, in particular to the gene sequence of triose phosphate isomerase of the monochamus alternatus and its cloning method. The gene encodes triose phosphate isomerase, which is an important material for studying the development of schistosomiasis vaccine and has important immunological significance. technical background [0002] Triosephosphate isomerase (Triosephosphate isomerase) is a glycolytic enzyme that widely exists in nature. It exists in many organisms such as plants, mammals, insects and fungi. It can catalyze dihydroxyacetone phosphate and 3-phosphoglycerol Reversible reaction between aldehydes. Triose phosphate isomerase plays an important role in glycolysis, fatty acid synthesis, gluconeogenesis, and pentose phosphate pathways. Studies have shown that triose phosphate isomerase is also an important material for the development of schistosomiasis vaccines, and has important Immunolo...

Claims

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Application Information

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IPC IPC(8): C12N15/61C12N9/90C12N15/10
CPCC12N9/90C12Y503/01001
Inventor 吴华俊林同蔡紫玲罗淋淋
Owner SOUTH CHINA AGRI UNIV
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