Hybridization-selected translation of Bombyx mori high-cysteine chorion proteins in Xenopus laevis oocytes.

Abstract

Xenopus laevis oocytes were injected with poly(A)+-mRNA isolated from chorionating follicular epithelium of the domesticated silk moth (Bombyx mori). On two-dimensional gel electrophoresis, the resultant translation products comigrated with authentic, secreted, chorion standards, demonstrating that the frog oocyte system synthesizes and correctly process virtually all major chorion components. A cDNA clone has been shown to contain sequences complementary to those of mRNAs encoding B mori high-cysteine (Hc) chorion proteins Hc6-Hc11. mRNAs were selected by hybridization to plasmid m5000 DNA bound to diazobenzyloxymethyl-cellulose and subsequently translated in X. laevis oocytes into forms that comigrated with authentic chorion standards. The selection of a distinct subset of Hc mRNAs under stringent hybridization conditions (70% formamide/0.2 M NaCl, 60 degrees C) suggests that they are encoded by related genes. This is consistent with the pattern obtained by hybridizing radioactive m5000 DNA to Southern blots prepared from EcoRI-cleaved B. mori chromosomal DNA.