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《Acta Universitis Medicinalis Nahui》 2002-01
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Screening triosephosphate transferase gene from the schistosomula cDNA library by using polymerase chain reaction

Tong Chang, Wang Xuelong, Shen Jijia (Dept of Microbiology and Parasitology,Anhui Medical Universtiy Hefei 230032)  
Objective The clone of triosephosphate transferase gene was screened from the schistosomula japonicum cDNA library by polymerase chain reaction in order to establish the foundation of subclone and expression. Methods A pair of primers were synthesized according to the open reading frame of TPM gene in Genebank, the frequency of TPM gene in the cDNA library was measured by using PCR. On the basis of this frequency, a proper dose of phage was inoculated into 96 hole cell culture plate and was amplified by bacteria culture. The positive clone was located by using PCR. After three times selective amplification the positive was identified and transformed into plasmid by self-cycled. A pGEM-T vector was constructed after the plasmid was sequenced. Results The TPM gene was screened from the cDNA library of schistosomula. The cloning vector of the TPM gene was successfully constructed. Conclusion The background of researching the role of TPM in schistosoma glycometabolism was established through this procedure.
【Fund】: 安徽省科技厅科研基金 (编号 :0 0 2 2 0 31);; 安徽省教育厅科研基金资助 (编号 :99ji0 1112 )
【CateGory Index】: R346
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