盐胁迫下盐穗木DNA聚合酶λ基因的克隆和表达分析  

Cloning and Expression Analysis of HcDNA pol λ Gene from Halostachys caspica under Salt Stress

作　　者：张冀[1] 杜驰[1] 张富春[1] ZHANG Ji DU Chi ZHANG Fu -chun(Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, Chin)

机构地区：[1]新疆大学生命科学与技术学院/新疆生物资源基因工程重点实验室,乌鲁木齐830046

出　　处：《新疆农业科学》2017年第2期361-370,共10页Xinjiang Agricultural Sciences

基　　金：新疆重点实验室专项资金"盐穗木盐胁迫响应的转录组研究及重要基因的功能鉴定"(2014KL001);国家"973"计划前期研究专项"新疆荒漠盐生植物耐盐的生理及分子机制"(2012CB722204)~~

摘　　要：【目的】开展盐胁迫下DNA损伤修复基因的表达研究,有助于揭示DNA损伤修复与盐生植物耐盐的相关性。【方法】根据盐胁迫下盐穗木转录组测序结果,利用RACE技术克隆获得了盐穗木DNA损伤修复基因HcDNA聚合酶λ(HcDNA polλ)基因,开放阅读框1 335 bp,编码444个氨基酸。【结果】保守结构域分析显示,HcDNA polλ基因属于DNA聚合酶X家族成员,系统进化分析显示HcDNA polλ为独立的分支,亚细胞定位于细胞核,无信号肽且不含跨膜区域的亲水蛋白。实时荧光定量PCR分析表明,随着NaCl胁迫浓度的增加,同化枝的HcDNA polλ基因的表达逐渐上调,在300 mmol/L NaCl处理7和14 d时,HcDNA polλ的表达分别增加3和5倍。最后在700 mmol/L NaCl处理14 d时,HcDNA polλ的表达增加20倍,达到峰值。【结论】HcDNA polλ基因的表达受盐胁迫的诱导。[ Objective] The expression of DNA damage repair gene under salt stress is helpful to reveal the relationship between DNA damage repair and salt tolerance of halophyte. [ Method ] According to the transcriptome of Halostachys caspica under salt stress, a DNA damage repair gene DNA polymerase lambda （HcDNA pol λ ） was cloned from Halostachys caspica by RACE. [ Result ] Sequence analysis indicated that HcDNA pol λ contained an open reading frame of 1,335 bp, which encodes 444 amino acids. Conserved domain analysis showed that HcDNA pol λ was the number of DNA polymerase X family, and phylogenetic tree analysis indicated that HcDNA pol λ was an independent branch, which was an stable hydrophilic proteins sub - celled nuclear. Real time fluorescent quantitative PCR analysis showed that the expression of HcDNA pol λ of assimilating branches gradually up -regulated with the increase of NaCl concentration. At 300 mmol/L NaCl treatments, the expression levels of HcDNA pol λ were increased 3 fold and 5 fold for 7 days and for 14 days, respectively. Finally, at 700 mmol/L NaCl treatment for 14 days, the expression levels of HcDNA pol λ increased 20 fold and reached the peak. [ Conclusion] The expression of HcDNA pol λ gene could be induced by salt stress.

关 键 词：盐穗木 HcDNA polλ基因 盐胁迫 基因表达 

分 类 号：S188[农业科学—农业基础科学]