摘要： 通过构建盐角草的液泡型Na+/H+反向转运蛋白SeNHX1基因的酵母表达载体,研究该基因对酵母耐盐性的影响。将SeNHX1基因与荧光报告基因GFP融合,分别构建酵母表达载体pYES2-SeNHX1-GFP、pYES2-SeNHX1和pYES2-GFP,转化酵母W303,诱导3个载体表达,并测定含有不同表达载体的酵母生长曲线。结果表明,荧光显微镜下观察到带有报告基因GFP的2个转化子W303(pYES2-SeNHX1-GFP)和W303(pYES2-GFP)均可发出绿色荧光,而没有报告基因的转化子W303(pYES2-SeNHX1)不发出荧光;在含有0.8 mol/L NaCl培养基中培养0~48 h内,W303(pYES2-SeNHX1-GFP)和W303(pYES2-SeNHX1)菌株前期生长比对照菌株W303(pYES2-GFP)慢,但在培养48 h后则大大超过对照菌株。表明所构建带有目的基因SeNHX1的酵母表达载体已表达,且证实SeNHX1基因可提高酵母的耐盐性。
关键词:
SeNHX1基因,
GFP基因,
酵母,
表达载体,
生长曲线,
荧光,
耐盐性
Abstract: In order to study effect of the gene SeNHX1 on yeast tolerance to salt, vector pYES2 - SeNHX1 -GFP、 pYES2 - SeNHX1 and pYES2 - GFP was constructed and transformed into yeast W303.Transformed W303 was cultured in CM -ura medium with D -gal which induced the vector expressing in Yeast1Bright green under fluorescence microscopewas observed in W303( pYES2 -SeNHX1 -GFP) and W303( pYES2 - GFP), but not in W303( pYES2 -SeNHX1).W303( pYES2 - SeNHX1 -GFP) and W303( pYES2 - SeNHX1) grew more slowly than W303( pYES2 - GFP) in 0- 48 h, it grew fast after 48 h1It indicated that the vector expressed correctly in yeast and gene SeNHX1 could enhance the tolerance to salt.
Key words:
SeNHX1 gene,
GFP gene,
Yeast,
Vector,
Growth curve,
Fluorescence,
Salt tolerance
中图分类号:
杨小玲, 季静, 王罡, 杨少辉, 孙建学, 张达威. SeNHX1与GFP融合基因在酵母中表达及其耐盐性表现[J]. 华北农学报, 2008, 23(4): 60-64. doi: 10.7668/hbnxb.2008.04.014.
YANG Xiao-ling, JI Jing, WANG Gang, YANG Shao-hui, SUN Jian-xue, ZHANG Da-wei. Expressing of Fusion Gene of SeNHX1 and GFP and Its Tolerance to Salt in Yeast[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(4): 60-64. doi: 10.7668/hbnxb.2008.04.014.