南极适冷菌Psychrobacter sp.G冷激蛋白基因Csp2039的表达分析  

Expression Characteristics of Cold Shock Protein Gene Csp2039 of the Antarctic Psychrotrophic Bacterium Psychrobacter sp.G

作　　者：李阳[1,2] 车帅[1,2] 王桢[1,2] 林学政[1,2] 

机构地区：[1]国家海洋局第一海洋研究所,山东青岛266061 [2]国家海洋局海洋生物活性物质重点实验室,山东青岛266061

出　　处：《海洋科学进展》2016年第1期85-94,共10页Advances in Marine Science

基　　金：国家自然科学基金项目--南极适冷菌Psychrobacter sp.G温度与盐度胁迫下基因表达谱分析及冷/热激基因应答机制研究(41176174);南北极环境综合考察与评估专项--北极海域海洋生物和生态考察(CHINARE2015-03-05)

摘　　要：以南极适冷菌Psychrobacter sp.G为研究对象,利用qRT-PCR和Western blot技术从转录水平和翻译水平对冷激蛋白基因Csp2039在不同温度/盐度胁迫条件下的表达特征进行了研究。在转录水平上,qRT-PCR分析表明,冷激蛋白基因Csp2039的表达于6h时显著被低温(0℃,10℃)和高温(30℃)诱导;低盐(0)胁迫下,基因Csp2039的表达先被诱导后被抑制,在盐度为15的胁迫下,基因Csp2039的表达均被诱导,高盐(90,120)时基因Csp2039的表达在6h时显著被诱导;在温度和盐度协同胁迫条件下,温度为0℃时,无论盐度高低,基因Csp2039的表达均显著上升,而高温(30℃)时,除2h被低盐诱导以外,其余条件下均显著被抑制。在翻译水平上,Western blot分析表明,冷激蛋白Csp2039的表达被低温(0℃,10℃)显著诱导,但高温(30℃)对其抑制并不明显;在低盐(0,15)胁迫下,Csp2039蛋白的表达均被诱导,高盐(90)胁迫下,Csp2039蛋白的表达先被抑制,然后逐渐升高;在温度和盐度协同胁迫条件下,低温(0℃)时,当盐度为15时,Csp2039蛋白的表达被诱导,盐度为90时,Csp2039蛋白的表达量被显著抑制;而高温(30℃)时,该蛋白在盐度15时6h的表达量最大,盐度90时于6h其表达被显著抑制。比较分析表明,在转录水平上基因Csp2039表达量的变化更容易受温度影响,而在翻译水平上盐度对Csp2039蛋白的影响作用大于温度;基因Csp2039在翻译水平上对温度/盐度胁迫的应答时间要迟于转录水平的。To clarify the adaption mechanism of microorganisms in the severe environment of Antarctica, a cold shock protein gene Csp2039 was cloned from the Antarctic psychrotrophic bacterium Psychrobacter sp. G. qRT-PCR and Western blot were used to investigate the expression characteristics of Csp2039 un- der different temperature/salinity stresses at the level of both transcriptional and translational. At the transcriptional level, qRT PCR showed that the expression of Csp2039 gene was significantly affected by the temperature shift and remarkably enhanced at 6 h by low temperature （0℃, 10℃）, and high temper- ature （30 ℃）. Under low salinity （0） stress, the expression of Csp2039 gene was first induced, and then was suppressed; at the salinity of 15, the gene expressions were all increased at different times （2, 6 and 12 h）; At high salinity （90, t20）, the expression was enhanced at 6 h. Under the combined temperature and salinity stress, the expression of Csp2039 gene was significantly increased at low temperature （0 ℃） regardless of salinity; on the contrary, the expression of Csp2039 gene was inhibited by high temperature （30℃） except at 2 h. In the translational pattern, Western blot indicated that the expression of Csp2039 protein was remarkably enhanced by low temperature （0℃, 10℃）, and obviously inhibited by high tem perature （30 ℃）. Under the low salinity （0, 15） stress, the expression of Csp2039 protein was enhanced; at the high salinity （90, 120）, the expression of Csp2039 gene was gradually increased after being sup- pressed. Under the combined temperature and salinity stresses, at low temperature （0℃）, the expression of Csp2039 protein was improved under the salinity of 15 while decreased under the salinity of 90; howev- er, at the high temperature （30℃）, it had a high expression at 6 h under the salinity of 15 while a low ex- pression at 6 h under the salinity of 90. In summary, the expression of Csp2039 gene was more likely to be affect

关 键 词：PSYCHROBACTER 冷激蛋白基因Csp2039 QRT-PCR WESTERN BLOT 表达分析 

分 类 号：Q751[生物学—分子生物学]