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利用卵壳cDNA文库对家蚕颗粒酶B突变进行分子分析。

Molecular analysis of the GrB mutation in Bombyx mori through the use of chorion cDNA library.

作者信息

Iatrou K, Tsitilou S G, Goldsmith M R, Kafatos F C

出版信息

Cell. 1980 Jul;20(3):659-69. doi: 10.1016/0092-8674(80)90312-8.

Abstract

To study the molecular basis of the GrB mutation, which prevents the synthesis of many stage-specific chorion proteins, a cDNA library has been constructed from wild-type chorion mRNA of Bombyx mori strain 703. By differential screening of the library with +/+ and B/B mRNAs, under appropriately stringent conditions to minimize cross hybridizations of related chorion sequences, we have selected several distinct clones corresponding to RNA sequences which are affected by the mutation (that is, are represented only in +/+ mRNA) or are unaffected (that is, are represented in both +/+ and B/B mRNAs). We show by Southern analysis that, whereas unaffected gene sequences are represented in both +/+ and B/B chromosomal DNA, affected sequences have been deleted from B/B DNA. The organization and regulation of developmental stage-specific chorion genes are discussed in light of these findings and the known effects of GrB on stage-specific protein synthesis.

摘要

为了研究GrB突变的分子基础,该突变阻止了许多阶段特异性绒毛膜蛋白的合成,我们从家蚕703品系的野生型绒毛膜mRNA构建了一个cDNA文库。通过用+/+和B/B mRNA对文库进行差异筛选,在适当严格的条件下以尽量减少相关绒毛膜序列的交叉杂交,我们选择了几个不同的克隆,它们对应于受突变影响的RNA序列(即在+/+ mRNA中才有)或未受影响的序列(即在+/+和B/B mRNA中都有)。我们通过Southern分析表明,未受影响的基因序列在+/+和B/B染色体DNA中都有,而受影响的序列已从B/B DNA中缺失。根据这些发现以及GrB对阶段特异性蛋白质合成的已知影响,我们讨论了发育阶段特异性绒毛膜基因的组织和调控。

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