北京师范大学学报(自然科学版)
北京師範大學學報(自然科學版)
북경사범대학학보(자연과학판)
JOURNAL OF BEIJING NORMAL UNIVERSITY (NATURAL SCIENCE)
2001年
2期
250-254
,共5页
脱水蛋白%表达载体%PCR%基因表达%热溶性分析
脫水蛋白%錶達載體%PCR%基因錶達%熱溶性分析
탈수단백%표체재체%PCR%기인표체%열용성분석
以克隆载体pTZ-19R-dhn1（ZM）为基础，构建脱水蛋白DHN1表达载体pBV221-dhn1.采用PCR技术从克隆载体pTZ-19R-dhn1（ZM）上扩增dhn1片段，并引入NcoⅠ/BamHⅠ酶切位点，然后与pBV221原核表达载体连接，得到pBV221-dhn1表达载体.阳性克隆经PCR和NcoⅠ/BamHⅠ酶切检测都得到516bp的 dhn1片段，且序列正确.表达载体pBV221-dhn1转化宿主菌后能够表达产生相对分子质量为22×103的DHN1，该蛋白具有高温可溶性.以上结果表明，本实验得到了高效表达的脱水蛋白DHN1表达载体pBV221-dhn1.
以剋隆載體pTZ-19R-dhn1（ZM）為基礎，構建脫水蛋白DHN1錶達載體pBV221-dhn1.採用PCR技術從剋隆載體pTZ-19R-dhn1（ZM）上擴增dhn1片段，併引入NcoⅠ/BamHⅠ酶切位點，然後與pBV221原覈錶達載體連接，得到pBV221-dhn1錶達載體.暘性剋隆經PCR和NcoⅠ/BamHⅠ酶切檢測都得到516bp的 dhn1片段，且序列正確.錶達載體pBV221-dhn1轉化宿主菌後能夠錶達產生相對分子質量為22×103的DHN1，該蛋白具有高溫可溶性.以上結果錶明，本實驗得到瞭高效錶達的脫水蛋白DHN1錶達載體pBV221-dhn1.
이극륭재체pTZ-19R-dhn1（ZM）위기출，구건탈수단백DHN1표체재체pBV221-dhn1.채용PCR기술종극륭재체pTZ-19R-dhn1（ZM）상확증dhn1편단，병인입NcoⅠ/BamHⅠ매절위점，연후여pBV221원핵표체재체련접，득도pBV221-dhn1표체재체.양성극륭경PCR화NcoⅠ/BamHⅠ매절검측도득도516bp적 dhn1편단，차서렬정학.표체재체pBV221-dhn1전화숙주균후능구표체산생상대분자질량위22×103적DHN1，해단백구유고온가용성.이상결과표명，본실험득도료고효표체적탈수단백DHN1표체재체pBV221-dhn1.
pBV221-dhn1, an expression vector for dehydrin DHN1, is constructed from its cloning vector pTZ-19R-dhn1 (ZM) and its gene expression is studied .The dhn1 fragment with NocⅠ/BamHⅠrestriction sites is generated by PCR through cloning vector pTZ-19R-dhn1 (ZM), and cloned into the expression vector pBV221 with T4 ligase to produce the expression vector pBV221-dhn1. After transformation, positive clones are analyzed by PCR amplification and restriction enzyme analysis, and a 516bp fragment with the same size and sequence of predicted dhn1 is obtained. Expression study shows that a specific protein of 22×103 with the characteristics of heat resistance is produced after gene transformation. Taken together, a highly expressed dehydrin expression vector pBV221-dhn1 is obtained.
