金发草<em>LEA 3 </em>基因两个剪接体转化酿酒酵母的抗非生物胁迫功能分析

中国生物工程杂志

2012, Vol. 32

Issue (08): 30-35

研究报告

金发草LEA 3 基因两个剪接体转化酿酒酵母的抗非生物胁迫功能分析

李锐¹, 王文国², 范林洪¹, 王胜华¹

1. 四川大学生命科学学院生物资源与生态环境教育部重点实验室成都 610064;
2. 农业部沼气科学研究所生物质能技术研究中心成都 610041

Abiotic Stress Tolerance Analysis Two Alternatively Spliced Isoforms of LEA3 Gene from Pogonatherum paniceum in Yeast

LI Rui¹, WANG Wen-guo², FAN Lin-hong¹, WANG Sheng-hua ¹

1. College of Life Sciences, Sichuan University, Department of Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Chengdu 610064, China;
2. Research Centre of Biomass Energy Technology, Biogas Institute of Ministry of Agriculture,Chengdu 610041, China

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摘要： 对金发草(Pogonatherum paniceum)第3组LEA蛋白( PpLEA3 )基因两个剪接体进行分析,并利用酿酒酵母表达系统分析两个剪接体在不同非生物胁迫的响应差异。以 PpLEA3 基因两个剪接体( PpLEA3.a 和PpLEA3.b )的重组载体pMD19-T- PpLEA3.a 和pMD19-T- PpLEA3.b 为模板,PCR法构建酵母表达载体pYES2- PpLEA3.a 和pYES2- PpLEA3.b ,并转化酿酒酵母细胞得到重组菌INV- PpLEA3.a 和INV- PpLEA3.b 。通过比较重组菌和对照菌(转空载体pYES2)在NaCl、NaHCO₃、低温、干旱、UV胁迫下的恢复生长状况,结果表明两种重组菌胁迫后的生长情况明显好于对照菌,两个剪接体对非生物胁迫抵抗力的大小为: PpLEA3.a > PpLEA3.b 。两个剪接体在核酸序列上的差异导致了在蛋白亲水性和结构上的差异,最终导致了在抗逆能力方面的差异。

关键词： LEA3基因; 金发草; 可变剪接; 转基因酵母; 非生物胁迫

Abstract: It was to analyze the sequence of Pogonatherum paniceum group 3 LEA proteins alternatively spliced isoforms, and to detect the abiotic stresses tolerance of PpLEA3 spliced isoforms. The two spliced isoforms of PpLEA3 gene were amplified by PCR reaction using the plasmids pMD19-T- PpLEA3.a and pMD19-T- PpLEA3.b as the templates, respectively. The yeast expression plasmid of pYES2- PpLEA3.a and pYES2- PpLEA3.b was constructed and then transformed into yeast to create recombinant INV- PpLEA3.a and INV- PpLEA3.b . Stress tolerance tests showed that LEA3 yeast transformants exhibited a higher survival rates than the control transformants under salt (NaCl), NaHCO₃, freezing, drought and ultraviolet radiation. PpLEA3.a has stronger abiotic stresses tolerance than PpLEA3.b . The nucleic acid sequence of two splicing isoforms have different protein hydrophilicity and structure which leading to differences in the stress tolerance.

Key words: LEA3gene Pogonatherum paniceum Alternative splicing Transgenic yeast Abiotic stress

收稿日期: 2012-03-13 出版日期: 2012-08-25

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通讯作者: 王胜华 E-mail: shwang200@yahoo.com.cn

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引用本文:

李锐, 王文国, 范林洪, 王胜华. 金发草LEA 3 基因两个剪接体转化酿酒酵母的抗非生物胁迫功能分析[J]. 中国生物工程杂志, 2012, 32(08): 30-35.

LI Rui, WANG Wen-guo, FAN Lin-hong, WANG Sheng-hua. Abiotic Stress Tolerance Analysis Two Alternatively Spliced Isoforms of LEA3 Gene from Pogonatherum paniceum in Yeast. China Biotechnology, 2012, 32(08): 30-35.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I08/30

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