Shirk P D, Perera O P
Center for Medical, Agricultural and Veterinary Entomology, U. S. Department of Agriculture, Gainesville, Florida, USA.
Arch Insect Biochem Physiol. 1998;39(3):98-108. doi: 10.1002/(SICI)1520-6327(1998)39:3<98::AID-ARCH2>3.0.CO;2-K.
The 5' region of YP2 cDNA, a follicular epithelium yolk protein subunit in the moth, Plodia interpunctella, shows that the polypeptide contains an extended internal coding region. Partial cDNA clones for YP2 were isolated from a pharate adult female ovarian cDNA expression library in Lambda Zap II by screening with antigen selected YP2 antiserum. The 5' sequence of the YP2 transcript was determined by 5' RACE PCR of ovarian mRNA using YP2 sequence-specific nested primers. The combined cDNA and 5' RACE sequencing showed the YP2 transcript to be 1971 bp in length up to the poly(A) tail with a single open reading frame for a predicted polypeptide of 616 amino acids. Northern analysis showed a single YP2 transcript to be present in ovarian RNA that was approximately 2 kb in length. The predicted amino acid sequence for YP2 from P. interpunctella is most closely related to egg specific protein (ESP) from Bombyx mori and the partial YP2 sequence from Galleria mellonella. YP2 from P. interpunctella also is similar to vertebrate lipases and contains a conserved lipid binding region. However, the 5' coding region of YP2 from P. interpunctella contains an in-frame insert of approximately 438 bp that had replaced an approximately 270-bp region as compared with ESP from B. mori and YP2 of G. mellonella. This suggests that the insert occurred by a recombinational event internal to the YP2 structural gene of P. interpunctella.
印度谷螟(Plodia interpunctella)卵泡上皮卵黄蛋白亚基YP2 cDNA的5'区域显示,该多肽包含一个延伸的内部编码区。通过用抗原筛选得到的YP2抗血清,从Lambda Zap II载体中的预蛹期成年雌性卵巢cDNA表达文库中分离出YP2的部分cDNA克隆。利用YP2序列特异性巢式引物,通过卵巢mRNA的5' RACE PCR确定YP2转录本的5'序列。cDNA和5' RACE测序结果表明,YP2转录本长度达1971 bp直至poly(A)尾,有一个单一的开放阅读框,预测的多肽含616个氨基酸。Northern分析显示,卵巢RNA中存在一条约2 kb长的单一YP2转录本。印度谷螟YP2的预测氨基酸序列与家蚕的卵特异性蛋白(ESP)以及大蜡螟的部分YP2序列关系最为密切。印度谷螟的YP2也与脊椎动物脂肪酶相似,含有一个保守的脂质结合区域。然而,与家蚕的ESP和大蜡螟的YP2相比,印度谷螟YP2的5'编码区含有一个约438 bp的框内插入片段,该片段取代了一个约270 bp的区域。这表明该插入是由印度谷螟YP2结构基因内部的重组事件导致的。