Multiple gamma-crystallins of the mouse lens: fractionation of mRNAs by cDNA cloning.

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RESUMO

cDNAs made from polyadenylylated RNAs of the mouse lens were cloned by the G.C tailing procedure in the bacterial plasmid pBR322. Four recombinant DNAs containing gamma-crystallin sequences were identified by hybrid selection and translation. Sequence analysis of the in vivo-labeled gamma-crystallin polypeptides that cofocused isoelectrically with the hybrid-selected translation products established that the four cloned cDNAs were derived from mRNAs encoding gamma-crystallin polypeptides with similar NH2 termini. The cDNA clones had different restriction maps and could discriminate among the different gamma-crystallin mRNAs under stringent hybridization conditions. Under relaxed hybridization conditions, the cDNA clones cross-hybridized with all gamma-crystallin mRNAs, and even slightly with beta-crystallin mRNAs, as judged by in vitro translation. RNA blot hybridization showed that the mouse lens gamma-crystallin mRNAs are 840 +/- 100 nucleotides long. These data indicate that there are at least four similar gamma-crystallin mRNAs and suggest (but do not establish) the existence of a closely related family of gamma-crystallin genes.

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