遗传
遺傳
유전
HEREDITAS(BEIJING)
2015年
2期
183-191
,共9页
吴日%马超%李晓丹%段会坤%姬艳丽%王宇%姜苹哲%王海松%屠培培%李淼%尼钢钢%马百成%李明刚
吳日%馬超%李曉丹%段會坤%姬豔麗%王宇%薑蘋哲%王海鬆%屠培培%李淼%尼鋼鋼%馬百成%李明剛
오일%마초%리효단%단회곤%희염려%왕우%강평철%왕해송%도배배%리묘%니강강%마백성%리명강
2型糖尿病%胰高血糖素样肽-1(GLP-1)%长效促胰岛素降糖酵母(LHY)%重组酿酒酵母%糖尿病小鼠模型
2型糖尿病%胰高血糖素樣肽-1(GLP-1)%長效促胰島素降糖酵母(LHY)%重組釀酒酵母%糖尿病小鼠模型
2형당뇨병%이고혈당소양태-1(GLP-1)%장효촉이도소강당효모(LHY)%중조양주효모%당뇨병소서모형
type 2 diabetes mellitus%glucagon-like peptide-1(GLP-1)%long-acting GLP-1 hypoglycemic yeast (LHY)%recombinant yeast%mouse model of type 2 diabetes mellitus
益生菌生物药物是指通过口服表达药用多肽(蛋白)的重组益生菌活细胞达到治疗疾病的新型口服给药系统。为了构建一种能有效防治2型糖尿病的酵母生物药物，文章首先构建了酿酒酵母(S.cerevisiae)整合型表达载体pNK1-PGK，并且通过绿色荧光蛋白(GFP)证明其表达功能正常，利用该载体将10×GLP-1(Glucagon-like peptide-1)基因转化到酿酒酵母INVSc1中，通过营养缺陷型和Western blotting成功筛选出表达10×GLP-1的长效促胰岛素降糖酵母(Long-acting GLP-1 hypoglycemic yeast, LHY)。该酵母生长迅速，外源基因10×GLP-1表达稳定，表达量达到1.56 mg/g细胞湿重。通过链脲佐菌素和高脂高糖饮食联合诱导的方法构建了2型糖尿病小鼠模型，用LHY对其进行口服灌胃治疗，证明LHY具有较好疗效，明显降低血糖水平。
益生菌生物藥物是指通過口服錶達藥用多肽(蛋白)的重組益生菌活細胞達到治療疾病的新型口服給藥繫統。為瞭構建一種能有效防治2型糖尿病的酵母生物藥物，文章首先構建瞭釀酒酵母(S.cerevisiae)整閤型錶達載體pNK1-PGK，併且通過綠色熒光蛋白(GFP)證明其錶達功能正常，利用該載體將10×GLP-1(Glucagon-like peptide-1)基因轉化到釀酒酵母INVSc1中，通過營養缺陷型和Western blotting成功篩選齣錶達10×GLP-1的長效促胰島素降糖酵母(Long-acting GLP-1 hypoglycemic yeast, LHY)。該酵母生長迅速，外源基因10×GLP-1錶達穩定，錶達量達到1.56 mg/g細胞濕重。通過鏈脲佐菌素和高脂高糖飲食聯閤誘導的方法構建瞭2型糖尿病小鼠模型，用LHY對其進行口服灌胃治療，證明LHY具有較好療效，明顯降低血糖水平。
익생균생물약물시지통과구복표체약용다태(단백)적중조익생균활세포체도치료질병적신형구복급약계통。위료구건일충능유효방치2형당뇨병적효모생물약물，문장수선구건료양주효모(S.cerevisiae)정합형표체재체pNK1-PGK，병차통과록색형광단백(GFP)증명기표체공능정상，이용해재체장10×GLP-1(Glucagon-like peptide-1)기인전화도양주효모INVSc1중，통과영양결함형화Western blotting성공사선출표체10×GLP-1적장효촉이도소강당효모(Long-acting GLP-1 hypoglycemic yeast, LHY)。해효모생장신속，외원기인10×GLP-1표체은정，표체량체도1.56 mg/g세포습중。통과련뇨좌균소화고지고당음식연합유도적방법구건료2형당뇨병소서모형，용LHY대기진행구복관위치료，증명LHY구유교호료효，명현강저혈당수평。
Probiotics, i.e., bacteria expressing therapeutic peptides (protein), are used as a new type of orally ad-ministrated biologic drugs to treat diseases. To develop yeast strains which could effectively prevent and treat type 2 diabetes mellitus, we firstly constructed the yeast integrating plasmid pNK1-PGK which could successfully express green fluorescent protein (GFP) in Saccharomyces cerevisiae. The gene encoding ten tandem repeats of glucagon-like peptide-1(10×GLP-1) was cloned into the vector pNK1-PGK and the resulting plasmids were then transformed into the S. cerevisiae INVSc1. The long-acting GLP-1 hypoglycemic yeast (LHY) which grows rapidly and expresses 10×GLP-1 stably was selected by nutrition screening and Western blotting. The amount of 10×GLP-1 produced by LHY reached 1.56 mg per gram of wet cells. Moreover, the oral administration of LHY significantly reduced blood glucose level in type 2 diabetic mice induced by streptozotocin plus high fat and high sugar diet.
