摘要
利用PCR扩增及PCR测序在显微注射法产生的转基因小鼠中发现,整合在小鼠染色体上的肌球蛋白轻链2启动子(myosinlightchain2promoter,MLC2)-糜酶(chymase)外源融合基因存在两种形式,一种为全长的融合基因,另一种在糜酶结构基因的第一内含子中缺失了213bp的序列。RT-PCR结果表明,缺失了部分内含子序列的外源融合基因不能在转基因小鼠心脏中表达.而全长的外源融合基因则能较高水平地表达,竞争性PCR定量实验表明在200ng心脏总RNA反转录产物中约含5.05(±1.38)×106个糜酶cDNA分子。上述结果表明,糜酶结构基因的第一内含子可能对MLC2-糜酶基因的表达具有调控作用。
PCR amplification and PCR sequencing were used to detect the integration of myosin light chain 2 promoter (MLC2) chymase fusion gene in the chromosome of transgenic mice produced by microinjection.It showed that there were two types of integration,one being the full length heterologous fusion gene,and the other including a 213 bp deletion in the first intron of the chymase structural gene.The RT PCR and competitive PCR results showed that the heterologous fusion gene including an intron sequence deletion could not express in the heart of the transgenic mice,but the full length heterologous fusion gene could express at high level and 5 05(±1 38)×10 6 ( n =3) chymase cDNA molecules were detected in 200 ng of heart total RNA reverse transcription.All results indicated that there might be some regulatory elements in the deleted sequence,which could affect the expression of MLC 2 chymase fusion gene in the transgenic mice significantly.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
1999年第5期704-708,共5页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金!资助项目 ( 3 95 70 2 97)
