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地中海实蝇(Ceratitis capitata)的绒毛膜基因。II. 通过对卵巢文库进行差异筛选获得的三个新cDNA克隆的特征分析。

The chorion genes of the medfly, Ceratitis capitata. II. Characterization of three novel cDNA clones obtained by differential screening of an ovarian library.

作者信息

Tolias P P, Konsolaki M, Komitopoulou K, Kafatos F C

机构信息

Department of Cellular and Developmental Biology, Harvard University, Cambridge, Massachusetts 02138.

出版信息

Dev Biol. 1990 Jul;140(1):105-12. doi: 10.1016/0012-1606(90)90057-p.

Abstract

We have isolated three new chorion cDNA clones from a Ceratitis capitata ovarian library. Their isolation was accomplished by differential screening of the library using as probes 32P-labeled poly(A)+ mRNAs obtained from hand-staged medfly choriogenic versus prechoriogenic follicles. RNA blot hybridization analysis revealed that the genes corresponding to these clones have unique temporal profiles of mRNA accumulation, restricted to specific choriogenic stages. In addition, in vitro translation products encoded by these cDNAs approximately comigrated with polypeptides synthesized de novo in culture by choriogenic follicles. All three genes are located in regions of the medfly genome that are specifically amplified in female ovaries. DNA sequence analysis has revealed that one of these clones is derived from a homolog of the Drosophila melanogaster s38 chorion gene. It appears that, although D. melanogaster and C. capitata are separated by at least 120 million years of evolution, the mechanisms by which chorion genes are expressed and regulated during development have been well maintained. We suggest that the regulatory elements controlling the expression of sex-specific (e.g., chorion) genes may be isolated and used to construct transgenic medfly strains from which females could be eliminated by negative selection; such strains could be used as part of an effort to control this agricultural pest.

摘要

我们从地中海实蝇卵巢文库中分离出了三个新的绒毛膜cDNA克隆。通过使用从手工分期的地中海实蝇成卵期与成卵前期卵泡中获得的32P标记的聚腺苷酸加尾mRNA作为探针,对文库进行差异筛选来完成它们的分离。RNA印迹杂交分析表明,与这些克隆对应的基因具有独特的mRNA积累时间模式,仅限于特定的成卵期阶段。此外,这些cDNA编码的体外翻译产物与成卵期卵泡在培养中重新合成的多肽大致共迁移。所有这三个基因都位于地中海实蝇基因组中在雌性卵巢中特异性扩增的区域。DNA序列分析表明,这些克隆之一源自黑腹果蝇s38绒毛膜基因的同源物。看来,尽管黑腹果蝇和地中海实蝇在进化上相隔至少1.2亿年,但绒毛膜基因在发育过程中表达和调控的机制仍得到了很好的保留。我们建议,可以分离出控制性别特异性(例如绒毛膜)基因表达的调控元件,并用于构建转基因地中海实蝇品系,通过阴性选择可以消除其中的雌性;这样的品系可作为控制这种农业害虫的一部分努力来使用。

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