金发草LEA3基因两个剪接体转化酿酒酵母的抗非生物胁迫功能分析  

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

作　　者：李锐[1] 王文国[2] 范林洪[1] 王胜华[1] 

机构地区：[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

出　　处：《中国生物工程杂志》2012年第8期30-35,共6页China Biotechnology

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。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 （NaC1）, 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.

关 键 词：LEA3基因 金发草 可变剪接 转基因酵母 非生物胁迫 

分 类 号：Q819[生物学—生物工程]