Construction of a cloned library of expressed embryonic gene sequences from Xenopus laevis.
Construction of a cloned library of expressed embryonic gene sequences from Xenopus laevis.
复制标题
非洲爪蟾表达胚胎基因序列克隆文库的构建。
DOI：
10.1016/0012-1606(80)90392-9
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发表时间：
1980
影响因子：
2.7
通讯作者：
I. Dawid
中科院分区：
文献类型：
--
作者：
M. Dworkin;I. Dawid
TotalXenopus laevispoly(A)+RNA from embryos at stage 10 (early gastrulae) and stage 41 (tadpoles) was copied into double-stranded cDNA, inserted into plasmid pBR322 by poly(dA:dT) tailing, and introduced intoEscherichia coli. Recombinant DNA libraries were obtained consisting of 5000 clones derived from stage 10 poly(A)+RNA and 12,500 clones from stage 41 poly(A)+RNA. We selected at random 860 clones for analysis by colony hybridization with [32P]cDNA prepared from stage 10 and stage 41 poly(A)+RNA populations. About 20% of the clones gave a detectable hybridization signal with the homologous probe. We estimate from reconstruction experiments that any clone containing a sequence present at a level of about 0.06% in the [32P]cDNA probe can be detected. Poly(A)+RNA from stage 10 embryos is dominated by a few very prominent sequences while stage 41 poly(A)+RNA is characterized by a larger array of less abundant species. The colony hybridization method allows us to evaluate the developmental behavior of a large number of abundant RNA sequences.
