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Method for preparing trypanosoma brucei leucyl tRNA synthetase activated protein

A technology of Trypanosoma brucei and active protein, which is applied in the field of preparation of active protein and can solve the problems of low expression efficiency

Inactive Publication Date: 2011-01-05
SHANGHAI JIAO TONG UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] After searching the literature of the prior art, it was found that Meng Xianfang, Shi Jing, Liu Xiaochun, etc. published a paper entitled "The Human Histidyl tRNA Synthetase Analogue" on pages 2407-2409 of "Chinese Journal of Pathophysiology", Volume 21, No. 12, 2005 Prokaryotic Expression and Preparation of Polyclonal Antibody", the article gives a method for expressing and purifying human histidyl tRNA synthetase analogues, but the expression efficiency of this method for Trypanosoma brucei leucyl tRNA synthetase is low

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  • Method for preparing trypanosoma brucei leucyl tRNA synthetase activated protein
  • Method for preparing trypanosoma brucei leucyl tRNA synthetase activated protein
  • Method for preparing trypanosoma brucei leucyl tRNA synthetase activated protein

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[0024] The present invention is further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific conditions in the following examples, usually according to conventional conditions, such as molecular cloning such as Sambrook: the laboratory manual sees the conditions described in the 1989 edition of New York Cold Spring Harbor Laboratory publishing house, or according to the manufacturer's suggestion condition.

[0025] Step 1, acquisition of the gene of T. brucei leucyl tRNA synthetase

[0026] (1) Get 20 μg of genomic cDNA of Trypanosoma brucei as a template, and the primers used for amplification are two primers upstream and downstream of PAGE purification, wherein,

[0027] Primer 1 sequence: GGCTGG CATATG TCGACTGTACGAC;

[0028] Primer 2 sequence: TTC GGAT...

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Abstract

The invention relates to a method for preparing trypanosoma brucei leucyl tRNA synthetase activated protein in the technical field of biology. The method comprises the following steps of: designing a specific primer and amplifying a trypanosoma brucei genome; recovering cut glue to obtain a trypanosoma brucei leucyl tRNA synthetase gene; connecting the gene obtained in the last step into a cloning vector to construct a gene cloning plasmid; subcloning the cloning plasmid into an expression vector, identifying, and converting into escherichia coli; cultivating the converted escherichia coli monoclone and inducing; and purifying expressed fusion protein to obtain the trypanosoma brucei leucyl tRNA synthetase activated protein. The leucyl tRNA synthetase activated protein can be used as a target for drug screening and is widely applied to the field of drug screening.

Description

technical field [0001] The invention relates to a method for preparing an active protein in the field of biotechnology, in particular to a method for preparing an active protein of trypanosome brucei leucyl tRNA synthetase. Background technique [0002] Tropical parasitic diseases are one of the important diseases that endanger human health in the world. It has affected hundreds of millions of people worldwide, caused a large number of deaths, and produced serious social consequences. African trypanosomiasis, also known as sleeping sickness, is named after the drowsiness caused by the pathogenic parasite Trypanosomiasis invading the central nervous system. So far, the four drugs marketed for the treatment of human African trypanosomiasis have several shortcomings. Drug toxicity, lack of parenteral administration and guaranteed supply, and the emergence of drug resistance are all problems that need to be addressed . [0003] In recent years, with the vigorous development of...

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Application Information

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IPC IPC(8): C12N9/00C12N15/70C12R1/19
Inventor 李大伟姚璎周虎臣
Owner SHANGHAI JIAO TONG UNIV
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