新的J超家族芋螺毒素的克隆与进化分析  

Cloning and Phylogenetic Analysis of Novel J-Superfamily Conotoxins

作　　者：吴传侠[1,2] 刘珠果[2] 孙婷[2] 李斌元[1] 戴秋云[2] 

机构地区：[1]南华大学药学与生命科学院,湖南衡阳421001 [2]军事医学科学院生物工程研究所,北京100071

出　　处：《生物技术通讯》2012年第3期364-368,共5页Letters in Biotechnology

基　　金：国家重点基础研究发展计划(2010CB529802);国家自然科学基金(81173035)

摘　　要：目的:从中国南海芋螺中克隆新的J超家族芋螺毒素,并进行序列和进化分析。方法:以芋螺毒腺管总RNA为模板,采用3′-RACE及巢式PCR的方法扩增J超家族芋螺毒素基因,并将得到的目的基因与pMD18-T载体连接并转化大肠杆菌DH5α,经测序比对后,获得新的J超家族毒素,利用软件BioEdit、ClustalX及Mega5.05进行进化分析。结果:获得12个新的J超家族芋螺毒素前体肽序列,其氨基酸组成具有新颖性,在进化树上与已报道的J超家族毒素处于不同的进化分支。结论:12个新的J超家族毒素与已报道的毒素序列之间的同源性较低,是J超家族新成员。Objective： To identify and analyze new J-superfamily conotoxins from six Conus species from the South China Sea. Methods： Total RNA was extracted from the venom duct of Conus species and was reverse transcribed into eDNA. The sequences of J-superfamily conotoxins were cloned by 3＇-rapid amplification of eDNA ends（RACE） and nest PCR method. The expected bands were then connected into the pMD18-T vector and transformed into E.coli DH5α competent cells. Applying the NCBI database, novel J-superfamily conotoxins were identified. The characterization and phylogenetic tree of novel J-superfamily conotoxins were analyzed using BioEdit, ClustalW and MegaS.05 software. Results： Twelve novel J-superfamily conotoxins with different amino acids residues were identified from six Conus species. The analysis of phylogenetic tree showed that these novel toxins located in the different branch. Conclusion： The novel J-superfamily conotoxins exhibit less homology to the reported, and belong to the new members of J-superfamily conotoxins.

关 键 词：芋螺毒素 J超家族 基因克隆 进化分析 

分 类 号：Q78[生物学—分子生物学] Q516