摘要
用ＰＣＲ方法从鲑精ＤＮＡ中分离出编码蛙鱼降钙素（ｓＣＴ）基因，并将其插入带有谷脱甘肽转移酶（ＧＳＴ）基因的融合表达载体ｐＧＥＸ－２Ｔ上，转入大肠杆菌ＪＭ１０９中，ＩＰＴＧ诱导表达。融合蛋白（ＧＳＴ－ｓＣＴ）表达量占菌体总蛋白量的３８％～４０％。用亲和层析方法纯化该融合蛋白，达８０％电泳纯，降钙素酶联免疫试验呈阳性反应。这为生产重组ｓＣＴ进而研究它的功能和应用打下了基础。
The DNA fragment encoding salmon calcitonin was separated from the salmon DNAwith PCR method.It was recombined into the fusion expression vector(pGEX-2T) whichincluded a Glutathion-S-Transferase(GST)gene and was transformed into E. coli JM109and then expression was induced with IPTG, The fusion protein(GST-sCT)accounted for38%～40%of the total cellular protein and was purified from the soluble expression products by glutathion sepharose 4B affinity chromatography. The purity of GST-sCT is about80%and it showed a positive immunological reaction in calcitonin enzyme immunoassay.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
1996年第6期423-428,共6页
Acta Academiae Medicinae Sinicae
关键词
鲑鱼
降钙素
基因
融合蛋白
表达
克隆
大肠杆菌
salmon calcitonin gene
fusion gene
inducible expression
affinity chromatography
