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印度谷螟中卵泡上皮卵黄蛋白亚基YP4的互补DNA

cDNA of YP4, a follicular epithelium yolk protein subunit, in the moth, Plodia interpunctella.

作者信息

Perera O P, Shirk P D

机构信息

Center for Medical, Agricultural and Veterinary Entomology, U. S. Department of Agriculture, Gainesville, Florida 32604, USA.

出版信息

Arch Insect Biochem Physiol. 1999;40(3):157-64. doi: 10.1002/(SICI)1520-6327(1999)40:3<157::AID-ARCH5>3.0.CO;2-W.

Abstract

YP4, a subunit of the follicular epithelium yolk protein in the moth, Plodia interpunctella, is produced in the follicle cells during vitellogenesis and after secretion is taken up into the oocyte and stored in the yolk spheres for utilization during embryogenesis. In order to identify the cDNA clones for YP4, a degenerate PCR primer was designed to six amino acid residues identified in the NH2-terminal sequence of mature YP4. The YP4 degenerate primer plus T7 reverse PCR primer produced a PCR product from a cDNA library for the majority of the YP4 coding sequence. Combined cDNA and 5' RACE sequencing showed the YP4 transcript to be 991 bp in length with a single open reading frame for a predicted polypeptide of 299 amino acids. Northern analysis showed a single YP4 transcript was present in ovarian RNA that was approximately 1 kb in length. The predicted amino acid sequence for YP4 from P. interpunctella was most closely related to the predicted YP4 protein from the moth, Galleria mellonella, and the spherulin 2a protein from the slime mold, Physarum polycephalum.

摘要

YP4是螟蛾(Plodia interpunctella)卵泡上皮卵黄蛋白的一个亚基,在卵黄发生过程中由卵泡细胞产生,分泌后被摄取到卵母细胞中,并储存在卵黄球中,以供胚胎发育期间利用。为了鉴定YP4的cDNA克隆,针对成熟YP4的NH2末端序列中确定的六个氨基酸残基设计了一个简并PCR引物。YP4简并引物加上T7反向PCR引物从cDNA文库中扩增出了大部分YP4编码序列的PCR产物。cDNA和5' RACE测序结果显示,YP4转录本长度为991 bp,有一个单一的开放阅读框,预测的多肽由299个氨基酸组成。Northern分析表明,在卵巢RNA中存在一个长度约为1 kb的单一YP4转录本。螟蛾YP4的预测氨基酸序列与大蜡螟(Galleria mellonella)预测的YP4蛋白以及黏菌多头绒泡菌(Physarum polycephalum)的球菌素2a蛋白关系最为密切。

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