| 摘要: |
| 用cDNA末端快速扩增方法克隆得到了一个2652bp的盐藻(Dunalilla salina)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(Chlamydomonas reinhardtii)、人、小麦(Triticum aestivum)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。 |
| 关键词: 盐藻(Dunalilla salina) hsp70 RACE 热休克 光诱导 |
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| 基金项目: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) |
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| Cloning of cytosolic hsp70cDNA in Dunalilla salina and the expression of hsp70 mRNA |
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| Abstract: |
| A full-length cDNA of the Dunalilla salina cytosolic hap70 gene was cloned and sequenced using RACE technique. Northern blot was performed to investigate the expression patterns of the hsp70 of the D. salina cells upon shifting a culture from 23℃ to 40℃ and from low-to high-light growth conditions, respectively. A cDNA of 2652 bp was obtained and deduced to 650 amino acids with two ATPase domains (LGGGTFDVS, FEVKSTA) and a peptide binding domain (aralarrlrtacerakrtlss). The identities of deduced amino acids to the cytosolic hsp70 from Chlamydomonas reinhardtii, human Triticum aestivum and Scerevisiae were 83%, 80%, 81.5% and 80.5%, respectively. The level of hsp70 mRNA at 40℃ for 90 min was 3-fold higher than that in light stress at the same time. In contrast, a slow increase of hsp70 mRNA was observed after light stress. These findings suggest that the expression of cytosolic hsp70 mRNA of D. salina can be induced by both heat or light stresses. |
| Key words: Dunalilla salina hsp70 RACE heat shock light stress |
