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[1]许天委,张晓楠,刘煜,等.海马齿小热激蛋白SpHSP18.1基因的克隆及盐胁迫表达分析[J].应用与环境生物学报,2015,21(05):882-886.[doi:10.3724/SP.J.1145.2015.04062]
 XU Tianwei,ZHANG Xiaonan,LIU Yu,et al.Molecular clone of Sesuvium portulacastrum small heat shock protein SpHSP18.1 gene and its expression profiling under salt stress[J].Chinese Journal of Applied & Environmental Biology,2015,21(05):882-886.[doi:10.3724/SP.J.1145.2015.04062]
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海马齿小热激蛋白SpHSP18.1基因的克隆及盐胁迫表达分析()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
21卷
期数:
2015年05期
页码:
882-886
栏目:
研究论文
出版日期:
2015-10-25

文章信息/Info

Title:
Molecular clone of Sesuvium portulacastrum small heat shock protein SpHSP18.1 gene and its expression profiling under salt stress
作者:
许天委 张晓楠 刘煜 叶绵源 张世杰 刘然 刘盛琨 张颖
1琼台师范高等专科学校数理系 海口 571127 2海南师范大学生命科学学院 海口 571158
Author(s):
XU Tianwei ZHANG Xiaonan LIU Yu YE Mianyuan ZHANG Shijie LIU Ran LIU Shenkun ZHANG Ying
1Department of Mathmatics and Physics, Qiongtai Teachers College, Haikou 571127, China 2College of Life Science, Hainan Normal University, Haikou 571158, China
关键词:
海马齿小分子热激蛋白分子克隆盐胁迫
Keywords:
Sesuvium portulacastrum small heat shock protein clone salt stress
分类号:
Q945.79 + Q78
DOI:
10.3724/SP.J.1145.2015.04062
文献标志码:
A
摘要:
为探究海马齿高盐环境耐受性的分子机制,根据海马齿cDNA文库序列设计引物,采用RACE(Rapid amplification of cDNA ends)技术克隆得到海马齿小分子热激蛋白SpHsp18.1基因序列. Blast分析表明,该cDNA序列(968 bp)含完整开放阅读框(ORF)全长为489 bp,编码162个氨基酸. 等电点(pI)为6.19,分子量(Mr)大小为18 605.90. SpHSP18.1编码氨基酸序列具有分子伴侣蛋白特有的ACD保守区域(Alpha crystallin domain),与拟南芥、西瓜等植物的细胞质I型小热激蛋白具较高同源性(> 75%). 将SpHsp18.1基因克隆到PUCm-T原核表达载体上,得到重组菌株,通过过量表达探究其耐热胁迫能力. 结果表明,在37 ℃下重组菌株与野生型菌株的生长曲线基本相似,但在高温(45 ℃)下,重组菌株较对照组菌株存活率有显著提高. 荧光实时定量PCR分析表明,在不同浓度海水培养条件下,海马齿根部均表达SpHsp18.1基因,且随着海水浓度的增加,该基因的表达量呈现上升的趋势. 说明SpHsp18.1基因在海马齿应答高盐胁迫过程中起到一定的作用.
Abstract:
For analyzing the molecular mechanism of Sesuvium portulacastrum under salt stress, the small hot shock protein gene Hsp18.1 (968 bp) was cloned by using RACE technique. The primers were designed based on the cDNA library sequences. The result showed that SpHsp18.1 cDNA coded 162 amino acids with the full length of 968 bp. The pI of the encoded protein was 6.19 and the Mr was 18 605.90. The amino acid sequence of SpHsp18.1 had the Alpha crystal in domain belonging to chaperonin family protein. Blast analysis found high homology (> 75%) with the cytoplasm type I small heat shock proteins of plants including Arabidopsis and watermelon. To verify the possible function of SpHsp18.1 protein, the PUCm-T vector was chosen and the recombinant SpHsp18.1 was overexpressed in Escherihia coli. The results showed that the growth of the transformed cells was similar to that of wild type at 37 oC, but the recombinant gene significantly improved cell viability in thermal stress conditions (45 oC). The results of real time PCR analysis showed expression of SpHsp18.1 gene on the roots of Sesuvium portulacastrum under salt stress in each treatment. At the same time, the gene expression quantity increased with the increase of the concentration of sea water. These results suggested that SpHsp18.1 protein may play a role in the response of Sesuvium portulacastrum to heavy salt stress.

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备注/Memo

备注/Memo:
国家自然科学基金项目(31360173)、海南省自然科学基金项目(312091)和海南师范大学2015年度“大学生创新创业训练计划”项目资助 Supported by the National Natural Science Foundation of China (31360173), the Hainan Natural Science Foundation (312091) and the College Students’ Innovation Entrepreneurship Training Program of Hainan Normal University (2015)
更新日期/Last Update: 2015-10-29