摘要
在相差显微镜下显微切割野生型黑腹果蝇唾液腺X染色体的1区片段,抽提该片段的DNA并与PGEM3Z质粒重组,然后转化到JM103 E.colt中,共切割12个染色体,抽提到大约10pgDNA,经重组、转化获得9个重组子。提取其中5个重组质粒分析其重组片段,2个约为0.6kb,另3个约为3.0kb。2个约为0.6kb重组片断用依赖DNA模板的SP6RNA聚合酶体外合成~3H标记的RNA,以此作探针进行染色体原位杂交定位,被克隆片段定位在果蝇唾液腺X染色体1E,1F位点上。本文结果表明重组到PGEM3Z质粒中染色体DNA可直接作为合成RNA探针的模板。
The DNA fragments microdissected from 1 region of the salivary gland X chromosome of D.melanogaster were cloned into PGEM3Z plasmid. From more than 10pg of chromosome DNA, 9 recombinent clones were obtained. The five recombinent clones were harvested and analysed.The two recombinent fragments are about 0.6 kb, the other three recombinent fragments are about 3.0 kb. The two RNA probes were synthesed with DNA dependent SP6 RNA polymerase translation system in vitro from the two recombinet clones containing fragements about 0.6 kb, and locateded at the region 1E and 1F of the X chromosome of D.melanogaster by hybridization in situ.The results showed that with this method, RNA can be synthesized in vitro directory from the recombinant clones which contain the DNA fragments from the D.melarogaster chromosome.
出处
《癌变·畸变·突变》
CAS
CSCD
1993年第1期5-8,共4页
Carcinogenesis,Teratogenesis & Mutagenesis
关键词
RNA探针
原位杂交
果蝇
DNA
Drosophila melanogaster
microdissection of chromosome
RNA probe
hybridization in situ
