Conotoxin RNA isolation and its cDNA synthesis / 中国海洋药物

Two methods of different RNA extractions from Conus venom tube and toxin gland of seven cone snail species were performed. ThermoScript~ TM RNase H~ - Transcriptase and AMV Transcriptase were used to synthesize cDNA separately. The cDNAs were used as templates to amplify conotoxin genes by PCR.The optimized RNA isolation and cDNA synthesis methods were obtained according to RT-PCR results, which would be the basis for cloning of new conotoxin genes.

Isolation and characterization of cDNA clones for α- and β-subunits of ovine luteinizing hormone.

A cDNA library of ovine pituitary DNA in plasmid pBR322 has been constructed by conventional methods with certain modifications. The library was screened using partial cDNAs for rat α-subunit and LHβ. We have isolated cDNA clones for ovine α- subunit and LHβ. The identification of these clones was confirmed by partial sequencing. The clones bear about 80% sequence homology with the respective rat cDNAs in the sequenced regions and hybridize with the rat clones in 5 X SSC at 55°C. The ovine LHβ clone has an insert of about 650 bp and selects an RNA of about 750 bases in a northern blot. The α-subunit cDNA clone has an insert of about 550 bp; it has two internal Pst I sites and thus shows restriction-based differences from rat α-subunit cDNA, which does not have any Pst I site.