盐藻胞浆hsp70 cDNA的克隆及其mRNA的诱导表达  

Cloning of cytosolic hsp70cDNA in Dunaliella salina and the expression of hsp70 mRNA

作　　者：姜国忠[1] 许培荣[1] 牛向丽[1] 王建民[1] 袁保梅[1] 薛乐勋[1] 

机构地区：[1]郑州大学医学院细胞实验室,河南郑州450052

出　　处：《海洋科学》2005年第5期43-49,共7页Marine Sciences

基　　金：国家自然科学基金项目(30270031);河南省重大科技攻关项目(0122032500);河南省杰出人才创新基金项目(0221001900)

摘　　要：用cDNA末端快速扩增方法克隆得到了一个2652bp的盐藻(Dunaliellasalina)胞浆hsp70cDNA全长,编码着650个氨基酸残基的多肽。推导的氨基酸序列有2个ATP酶结合位点和一个多肽结合区。由克隆得到的盐藻cDNA推导的氨基酸序列,经GenBankblast后与数据库中胞浆hsp70序列一致。与莱茵衣藻(Chlamydomonasreinhardtii)、人、小麦(Triticumaestivum)和啤酒酵母(Saccharomyces.cerevisiae)胞浆hsp70的序列一致性分别为83%、80%、81.5%和80.5%。Northern印迹显示,90min后,mRNA量在40℃热休克时是光诱导时的3倍。光诱导则使hsp70mRNA积累相对迟缓。结果表明,所克隆得到的hsp70基因蛋白产物定位在盐藻细胞浆中,光诱导和热休克均可以使hsp70mRNA水平明显增高,但以热休克为显著。A full-length cDNA of the Dunaliella salina cytosolic hsp70gene was cloned and sequenced using RACE technique.Northern blot was performed to investigate the expression patterns of the hsp70of the D.salina cells upon shifting a culture from23℃to40℃and fromlow-to high-light growth conditions,respectively.A cDNA of2652bp was obtained and deduced to650amino acids with two ATPase domains(LGGGTFDVS,FEVKSTA)and a peptide binding domain(ARALRRLRTACERAKRTLSS).The identities of deduced amino acids to the cytosolic hsp70from Chlamydomonas reinhardtii,human,Triticum aestivum and Scerevisiae were83%,80%,81.5%and80.5%,respecˉtively.The level of hsp70mRNA at40℃for90min was3-fold higher than that in light stress at the same time.In contrast,a slowincrease of hsp70mRNA was observed after light stress.These findings suggest that the expression of cyˉtosolic hsp70mRNA of D.salina can be induced by both heat or light stresses.

关 键 词：盐藻(Dunalilla salina) HSP70 RACE 热休克 光诱导 

分 类 号：Q78[生物学—分子生物学]