摘要
以粟酒裂殖酵母基因组DNA为模版,逆转录获得cDNA;以cDNA为模板,采用聚合酶链式反应(PCR)技术扩增,得到苹果酸通透酶(mae1)基因,将其与pMD-18T连接并测序.结果表明cDNA全长为1316 bp,编码438氨基酸,分子质量49ku,与已报道的mae1基因序列的同源性达到99%.mae1与整合型表达质粒pSH47连接后,用重组的表达质粒转化产朊假丝酵母,经PCR验证筛选阳性克隆子.通过对转化子的摇瓶筛选,发现重组子的降酸幅度均大于原始菌株,培养40 h降解了48.6%的L-苹果酸,降解率比原始菌株提高了9.8%.
Genome DNA from Schizosaccharomyces pombe was reverse transcribed as template to acquire cDNA,and mae1 was reproduced by PCR with cDNA template.PDM-T carrier was inserted and the full length of cDNA chain was detected as 1316 bp,which encoded 438 amino acid with the molecular mass of 49 ku.The sequence analysis result showed it had 99% similarity with the reported one.Mae1 was conjointed with integrated vector pSH47 and transformed into Candida utilis.The positive transformants were detected by PCR.The deacidification capacity of yeast recombinants was compared by shake flask selection,48.6% of malic acid was degraded by CU-6 in 40 hours,which was 9.8% higher than control.
出处
《福建农林大学学报（自然科学版）》
CSCD
北大核心
2012年第5期513-517,共5页
Journal of Fujian Agriculture and Forestry University:Natural Science Edition
基金
福建省科技厅资助项目(2009N2002)
福建省教育厅资助项目(JB10017)
