THE PREPARATION OF RNA PROBES WITH CLONING OF DNA FROM MICRODISSECTED D.MELANOGASTER CHROMOSOME FRAGMENTS AND ITS LOCALIZATION BY HYBRIDIZATION IN SITU
Cai Ping, Wang Jinying, Chen Tiehe, Xiang Zhenghua, Mong Lin, Wang Xinmin, Li Huaiyi Naval Medical Research Institute, Shanghai 200433 Department of Histology and Embryology, Department of Biology Second Military Medical University
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.