期刊论文

Activity Analysis and Secretory Expression of Human Alpha-defensin 5 in Saccharomyces cerevisiae

中文

生命科学研究

1007-7847

2017

21

6

471-476

10.16605/j.cnki.1007-7847.2017.06.001

16A098:湖南省教育厅重点项目 2017NK2311:湖南省科技厅重点项目 31672457:国家自然科学基金 31772865:国家自然科学基金 XCX17071:湖南农业大学大学生创新项目

本校为第一机构

人α-防御素5 (human α-defensin5,HD5)是人防御素α家族中发现的抑菌活性最高的多肽。为了探究HD5在酿酒酵母中分泌表达的可行性,首先利用PCR扩增获得酵母菌偏好的HD5核酸序列,构建酿酒酵母表达载体pVT102U/α-HD5,并将该重组质粒转入酿酒酵母S78中,通过营养缺陷筛选获得阳性转化菌株。然后,对重组菌进行发酵培养,取上清液纯化后通过tricine-SDS-PAGE和质谱检测表达产物。最后,利用琼脂扩散法检测表达的HD5的抑菌活性以及其对温度的耐受性,同时通过二倍稀释法检测表达的HD5对大肠杆菌、金黄色葡萄球菌以及沙门氏杆菌的最小完全抑制浓度。结果显示:表达的HD5对大肠杆菌、金黄色葡萄球菌以及沙门...

The feasibility of secretion expression of human α-defensin 5 （HD5）, which exhibits the highest antimicrobial activity in human α-defensin family, in Saccharomyces cerevisiae was explored. The DNA fragment containing HD5 coding sequence with biased codons of S. cerevisiae was amplified by PCR, and inserted into the shuttle plasmid pVT102U/α of S. cerevisiae. The recombinant vector named pVT102U/α-HD5 was transformed into S. cerevisiae S78. The positive recombinant clones were screened out under nutrient deficiency. After fermentation, the...