拟穴青蟹PDI保守区域的克隆与序列分析  

Cloning and Sequence Analysis of Scylla paramamosain Protein-disulfide Isomerase Conserved Domain

作　　者：刘元婧[1] 杨玉荣[2] 叶海辉[1] 黄辉洋[1] 王桂忠[1] 李少菁[1] 

机构地区：[1]厦门大学海洋与环境学院 [2]厦门大学生命科学学院,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2009年第6期895-899,共5页Journal of Xiamen University：Natural Science

基　　金：国家自然科学基金(40406030;40776084);福建省高等学校新世纪优秀人才支持计划资助

摘　　要：蛋白二硫键异构酶（PDI）是真核生物重要的多功能蛋白，其基因结构在虾蟹类尚未报道．从拟穴青蟹Scyllapa。ramamosain（Estampador，1949）脑组织中分离纯化总RNA，经逆转录得到cDNA第一条链，并以之为模板利用RACE（rapid—amplificationofcDNAends）技术，扩增出一条950bp的cDNA．将所得cDNA克隆到质粒载体pTZS7R／T，转化大肠杆菌DH5a细胞并对筛选的阳性克隆测序．通过与目前数据库中序列的比较，在此cDNA编码蛋白中发现其中i01个氨基酸与其他物种已知蛋白二硫键异构酶（PDI）中的PDI_a_PDI_a＇_C保守区域相似率在64％～79％之间．确定此cDNA编码拟穴青蟹PDI_a_PDI_a＇_C保守区域．The protein disulfide isomerase severs to be s multifunctional protein of great importance to the eukaryotes. The total RNAs isolated from the brain of Scylla paramamosainn （Estampador, 1949） were reverse transcribed to the first strands of eDNA. A eDNA fragment of 950 bp was amplified from the first strands of eDNA by RACE （rapid-amplification of eDNA ends） method. This fragment was inserted into plasmid pTZ57R/T for screening and DNA sequencing. Compared with current sequences in the database, a 101-an domain,which was found in the putative protein, had 64% -79 % homology with the PDI_a_PDI_a＇_C conserved domains of protein-disulfide isomerases （PDIs） in other species. This eDNA was supposed to encode Scylla paramamosain PDI_a_PDI _a＇_C conserved domain.

关 键 词：蛋白二硫键异构酶 PDI_a_PDI_a'_C保守区域 拟穴青蟹 基因克隆 序列分析 

分 类 号：Q959.223[生物学—动物学] Q785