盐胁迫下盐穗木DNA聚合酶λ基因的克隆和表达分析

发布时间：2018-08-02 10:03

【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。
[Abstract]:[objective] to study the expression of DNA damage repair genes under salt stress, which is helpful to reveal the correlation between DNA damage repair and salt tolerance in halophytes. [methods] according to the results of DNA transcription sequencing under salt stress, The HcDNA polymerase 位 (HcDNA pol 位) gene of DNA damage repair gene was cloned by RACE technique. The open reading frame 1 335 BP encodes 444 amino acids. [results] the conserved domain analysis showed that HcDNA pol 位 gene belonged to DNA polymerase X family. Phylogenetic analysis showed that HcDNA pol 位 was an independent branch, subcells located in the nucleus, no signal peptide and no transmembrane region of hydrophilic protein. Real-time fluorescence quantitative PCR analysis showed that the expression of HcDNA pol 位 gene increased gradually with the increase of NaCl stress concentration, and increased by 3 and 5 times at the 7th and 14th day after treatment with 300 mmol/L NaCl. At last, the expression of HcDNA pol 位 increased 20 times to the peak at 700 mmol/L NaCl treatment for 14 days. [conclusion] the expression of HcDNA pol 位 gene was induced by salt stress.
【作者单位】：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;
【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)~~
【分类号】：Q943.2

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