不同环境因子对海洋细菌Pseudoalteromonas issachenkoniiHZ引导糖基转移酶基因表达的影响  

The impact of environmental factors on marine bacteria Pseudoalteromonas issachenkonii HZ glycosyltransferase gene expression

作　　者：卢晓平[1] 许艳蕊[1] 范秋苹 郝鲁江[1] 

机构地区：[1]齐鲁工业大学山东省微生物工程重点实验室,山东济南250353

出　　处：《食品工业科技》2016年第22期212-216,共5页Science and Technology of Food Industry

基　　金：山东省自然科学基金(ZR2012CM019)

摘　　要：为了研究海洋细菌Pseudoalteromonas issachenkonii HZ所产多糖的生物合成基因簇,首先需要克隆到在多糖合成中起关键作用的引导糖基转移酶基因。通过设计引物,采用PCR技术从海洋细菌Pseudoalteromonas issachenkonii HZ成功克隆到该基因,并通过实时荧光定量PCR技术检测了温度、p H及海盐浓度对引导糖基转移酶基因表达量的影响。结果表明:低温有利于引导糖基转移酶的表达,随着温度的上升,引导糖基转移酶基因的表达量先增后减,在20 h时,引导糖基转移酶的表达量急剧上升,15℃和35℃引导糖基转移酶表达量分别是20℃表达量的5.2倍、5.8倍;海盐浓度为4.5%时引导糖基转移酶的表达量为海盐浓度3.5%的2.5倍,海盐浓度为5.5%时引导糖基转移酶的表达量为海盐浓度3.5%的2.0倍;10 h时p H为8、9的培养基中引导糖基转移酶的表达量分别为p H为7的1.64和1.67倍。该结果为探究菌体环境适应性机制提供了一定的基础。To study the biosynthesis gene cluster of the polysaccharide produced by marine bacterium Pseudoalteromonas issachenkonii HZ, and to clone the gene which plays a key role in the synthesis of polysaccharide.Glycosyltransferase gene was amplified by designing primers using PCR technology from marine bacteria Pseudoalteromonas issachenkonii HZ,and examined the effect of temperature, pH and salt concentration of glycosyltransferase gene expression by real-time fluorescence quantitative PCR.The results showed that the low temperature was in favor of the expression of the glycosyltransferase.With the increase of the temperature, the expression of the gene of the glycosyltransferase increased first and then decreased. It was showed that glycosyltransferase gene expression level increased rapidly,and the glycosyltransferase gene expression level of Pseudoalteromonas issachenkonii HZ grown at 15 and 35 ℃ were 5.2,5.8 times higher than grown at 20 ℃, respectively.4.5% salt concentration could increase the expression level of glycosyltransferase gene 2.5 times compared to that of 3.5% salt concentration, while 5.5% salt concentration glycosyltransferase gene expression level was 2 times compared to 3.5% salt concentration. The glycosyltransferase gene expression level of Pseudoalteromonas issachenkonii HZ grown at pH8 and 9 were 1.64, 1.67 times higher than grown at pHT, respectively.The research on glycosyltransferase gene of Pseudoalteromonas issachenkonii HZ laid a foundation for exploring the mechanism of adaptation to the environment.

关 键 词：海洋细菌Pseudoalteromonas issachenkonii HZ 引导糖基转移酶 实时荧光定量PCR 

分 类 号：TS201.1[轻工技术与工程—食品科学]