毒腺_____中华眼镜蛇；毒腺；cDNA表达文库；PCR

中华眼镜蛇毒腺cDNA表达文库的建立和分析

http://www.100md.com 《第四军医大学学报》 2004年第13期

毒腺,,中华眼镜蛇；毒腺；cDNA表达文库；PCR,0引言,1材料和方法,2结果,3讨论,【参考文献】

     Construction of cDNA library from venom of Chinese Cobra

    WAN Rong, SONG Jun, LIN Xu， LIN JianYin

    1Molecular Medicine Research Center, 2Department of Pathology, Fujian Medical University, Fuzhou 350004, China

    【Abstract】 AIM: To construct a high quality cDNA expression library from the venom of Chinese Cobra. METHODS: Total RNA and purified mRNA were extracted by using "singlestep method" and by chromatography on oligo (dT) cellulose. The cDNA was synthesized through reverse transcription. After cDNA size fractionation was purified by chromatography column, the double cDNA was ligated to pSPORT1 plasmid vector. RESULTS: The directional library was confirmed to be about 2×105 independent clones in which the percentage of recombinant clones was 97% and the whole length cDNA of CTX3 and PLA2 gene were obtained by PCR amplification. CONCLUSION: The constructed cDNA library can be used for further screening and cloning of new protease inhibitor gene in Chinese cobra.

    【Keywords】 naja atra; venom; cDNA expression library; PCR

    【摘要】目的: 为研究开发眼镜蛇毒药用基因工程产品，筛选克隆及表达相关药用功能基因 ......

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