杜氏盐藻14-3-3蛋白cDNA片段的克隆与分析  

Cloning and analysis of 14-3-3 protein cDNA fragment of Dunaliella salina

作　　者：姬翔[1] 王天云[1] 王亚峰[1] 刘红涛[1] 王建民[1] 薛乐勋[1] 

机构地区：[1]郑州大学细胞生物学研究室

出　　处：《郑州大学学报（医学版）》2006年第3期433-435,共3页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金资助项目30470030;30270031;河南省医学科技创新人才工程项目2001006

摘　　要：目的:克隆杜氏盐藻14-3-3蛋白的cDNA片段,并对其进行测序和序列分析。方法:根据衣藻、烟草、豌豆、拟南芥、西红柿等14-3-3蛋白的高度保守序列SVAYKNV、DSTLIMQ设计一对简并引物,采用RT PCR方法扩增杜氏盐藻14-3-3蛋白的cDNA片段。PCR产物克隆至T载体后转化大肠杆菌JM109,筛选阳性菌落,测序,BLAST进行同源性比对。结果:克隆获得531bp的cDNA片段,编码177个氨基酸(GenBankAN:AY965896)。同源性比较显示,序列与其他生物具有高度的同源性,其与衣藻、烟草、豌豆、拟南芥、西红柿、人等的同源性分别为:94%,84%,84%,83%,83%和80%。结论:克隆得到了杜氏盐藻14-3-3蛋白的cDNA片段。Aim ： To clone and identify 14-3-3 protein eDNA fragment from DunalieUa salina. Methods：A pair of degenerate primers was designed according to the conserved motifs of SVAYKNV and DSTLIMQ of homologous amino acid sequences, and total RNA of DunalieUa salina was extracted using TRIzol reagent. A eDNA fragment obtained through RTPCR was inserted into T-vector and then transformed into E. coil JM109. Several positive colonies were randomly selected and screened for sequencing, Homologous analysis of the deduced amino acid sequences was performed by BLAST and subsequently compared with those of GenBank. Result.s： The cloned eDNA sequence was 531bp, which encoded 177 amino acids（GenBank accession number： AY965896）. The sequence shared high identity with the following 14-3-3 proteins： Chlamydomonas reinhardtii 94% , Nicotiana tabacum 84% , Pisum sativum 84% , Arabidopsis thaliana 83% , tomato 83% , and human beings 80% , respectively. Conclusion： The cloned sequence is 14-3-3 cDNA fragment from DunalieUa salina.

关 键 词：杜氏盐藻 14—3—3蛋白 CDNA 简并引物 

分 类 号：Q781[生物学—分子生物学]