cDNA library construction of the venom duct of Conus caracteristicus Fischer native to Hainan
会议名称：《第九届中国生物毒素学术研讨会》
会议日期：2009年
学科分类：100405[医学-卫生毒理学] 1004[医学-公共卫生与预防医学(可授医学、理学学位)] 10[医学]
关 键 词：Conus caracteristlcus Fischer venom ducts cDNA library construction
摘 要：To conserve the precious conotoxin gene resource of Conus caracteristicus Fischer native to Hainan,a cDNA library of the venom duct of *** Fischer was constructed,which would provide the biomaterial for investigating and developing the original marine conopeptide *** RNA was extracted from the venom ducts of *** Fischer by Total RNA Isolation Kit,and the Primer Extention method was used to synthesized double-strand cDNA based on the SMART *** digestion with SfiⅠand size fractionation by CHROMASPIN-400 columns,cDNAs(500bp) were ligated to dephosphorylated pDNR-LIB vector digested by SfiⅠ.Then,the recombinant plasmids were transformed into *** DH5αthrough *** original library was generated through pooling the desired transformation mixtures *** were picked randomly from the library for screening the size of the cDNA inserts through Bacteria Liquid *** last step was the library *** original constructed cDNA library contained 3.1×10 independent *** titer of the cDNA library was about 3.01×10 cfu/mL,with the percentage of recombinant clones about 90%.The average size of the inserts was about 1 *** constructed cDNA library meets the requirement of a standard cDNA *** could be a strong basis for subsequent work of finding and researching some kinds of new conotoxins from Conus caracteristicus Fischer.