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5' coding region of the follicular epithelium yolk polypeptide 2 cDNA in the moth, Plodia interpunctella, contains an extended coding region

机译:蛾的卵泡上皮卵泡上皮蛋黄多肽2 cDNA的5'编码区包含一个扩展的编码区

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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 femaleovarian 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 alsois 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 comparedwith 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.
机译:YP2 cDNA的5'区是蛾中的卵泡上皮卵泡卵黄蛋白亚基,表明该多肽包含一个扩展的内部编码区。通过用抗原选择的YP2抗血清筛选,从Lambda Zap II中的一个成年雌性成年女性卵巢cDNA表达文库中分离出YP2的部分cDNA克隆。使用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。组合的cDNA和5'RACE测序表明,YP2转录物的长度为1971 bp,直至poly(A)尾部,并具有一个单一的开放阅读框,可预测616个氨基酸。 Northern分析显示卵巢RNA中存在单个YP2转录本,其长度约为2 kb。穿刺假单胞菌的YP2的预测氨基酸序列与家蚕的卵特异性蛋白(ESP)和马勒菌廊的部分YP2序列最相关。穿刺假单胞菌的YP2也类似于脊椎动物脂肪酶,并包含一个保守的脂质结合区。然而,来自穿刺假单胞菌的YP2的5'编码区含有大约438bp的框内插入物,与来自桑蚕假单胞菌的ESP和melonella的YP2的ESP相比,其已替换了大约270bp的区域。这表明该插入片段是由点状毕赤酵母的YP2结构基因内部的重组事件引起的。

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