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家蚕卵泡中的细胞特异性表达:一种主要卵壳蛋白、其mRNA和基因的发育特征

Cell-specific expression in the silkmoth follicle: developmental characterization of a major chorion protein, its mRNA and gene.

作者信息

Regier J C, Cole C, Leclerc R F

机构信息

Center for Agricultural Biotechnology, University of Maryland, College Park 20742.

出版信息

Dev Biol. 1993 Nov;160(1):236-45. doi: 10.1006/dbio.1993.1301.

Abstract

Choriogenesis (eggshell formation) within the silkmoth Antheraea polyphemus proceeds in parallel for the two major subpopulations of follicle cells, diverging only during the very late period when aeropyle crown surface structures form in one region but not in the other. Correlated with their appearance is the synthesis of a set of region-specific proteins. In this report, aeropyle crowns are physically isolated and their protein composition is shown to consist of those same region-specific proteins. A cDNA clone, called pcvl 16, has been selected and shown to encode a lamellar-forming, aeropyle crown-specific protein, probably of the previously described C3,4 group. These conclusions are based on hybrid-selected translation, Northern analysis, and sequence analysis. pcvl 16 was used to isolate two distinct cloned copies of the 16 gene. Both 16 genes are closely paired with another region-specific gene but the proximity of the two gene pairs to each other is uncertain. Non-region-specific chorion genes expressed at earlier times in choriogenesis surround the 16 gene pairs, suggesting that cis sequences necessary for regionalized expression may be closely linked to coding sequences. To test this hypothesis, 5'-flanking sequences from eight region-specific genes are compared and shown to share two oligonucleotide sequences. One is a known regulatory element found in virtually all moth and fly chorion genes examined. The other, located just upstream from the TATA box, is not found in non-regionally expressed chorion genes and, thus, is a candidate for specifying regional expression.

摘要

在多音天蚕蛾中,卵泡细胞的两个主要亚群的绒毛发生(卵壳形成)过程是并行的,仅在气室冠表面结构在一个区域形成而在另一个区域不形成的非常晚期才出现分化。与它们的出现相关的是一组区域特异性蛋白质的合成。在本报告中,气室冠被物理分离,其蛋白质组成显示为由那些相同的区域特异性蛋白质组成。一个名为pcvl 16的cDNA克隆已被筛选出来,并显示编码一种形成片状的、气室冠特异性蛋白质,可能属于先前描述的C3,4组。这些结论基于杂交选择翻译、Northern分析和序列分析。pcvl 16被用于分离16基因的两个不同的克隆拷贝。两个16基因都与另一个区域特异性基因紧密配对,但这两个基因对彼此的接近程度尚不确定。在绒毛发生早期表达的非区域特异性绒毛膜基因围绕着16基因对,这表明区域化表达所需的顺式序列可能与编码序列紧密相连。为了验证这一假设,对八个区域特异性基因的5'侧翼序列进行了比较,结果显示它们共享两个寡核苷酸序列。一个是在几乎所有已检测的蛾类和蝇类绒毛膜基因中都发现的已知调控元件。另一个位于TATA框上游,在非区域表达的绒毛膜基因中未发现,因此是指定区域表达的一个候选元件。

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