Lu Daguang, Li Xiangrui, Liu Xiaoxia, Zhang Qingwen
Department of Entomology, College of Agriculture and Biotechnology, China Agricultural University, Beijing 100094, China.
J Chem Ecol. 2007 Jul;33(7):1359-75. doi: 10.1007/s10886-007-9310-5.
Two putative odorant-binding proteins (OBPs) and one putative chemosensory protein (CSP) from females of the ant-like bethylid wasp, Scleroderma guani Xiao et Wu (Hymenoptera: Bethylidae), were identified and cloned. The putative OBPs and CSP were identified by nondenaturing polyacrylamide gel electrophoresis (native-PAGE). 3' rapid amplification of cDNA ends (3'RACE) was performed to obtain the sequences of the mature proteins by using degenerate primers designed from N-terminal sequences. Gene-specific primers for 5' rapid amplification of cDNA ends (5'RACE) were designed according to 3'RACE results and used in polymerase chain reaction (PCR) to obtain full-length sequences. The proteins (Sgua-OBP1, Sgua-OBP2, and Sgua-CSP1) encode 133, 142, and 129 amino acid-deduced sequences, respectively. Prediction of signal peptide sequences matches the N-terminal amino acid sequence of the isolated proteins. Database searches suggest that the Sgua-OBP1 and Sgua-OBP2 are homologs of OBPs from other insects, and Sgua-CSP1 shares a high level of identity with previously described CSPs.
从类蚁甲腹茧蜂（Scleroderma guani Xiao et Wu，膜翅目：甲腹茧蜂科）雌蜂中鉴定并克隆出两种假定的气味结合蛋白（OBP）和一种假定的化学感受蛋白（CSP）。通过非变性聚丙烯酰胺凝胶电泳（native-PAGE）鉴定假定的OBP和CSP。利用根据N端序列设计的简并引物进行3' cDNA末端快速扩增（3'RACE），以获得成熟蛋白的序列。根据3'RACE结果设计用于5' cDNA末端快速扩增（5'RACE）的基因特异性引物，并用于聚合酶链反应（PCR）以获得全长序列。这些蛋白质（Sgua-OBP1、Sgua-OBP2和Sgua-CSP1）分别编码133、142和129个氨基酸推导序列。信号肽序列预测与分离蛋白的N端氨基酸序列匹配。数据库搜索表明，Sgua-OBP1和Sgua-OBP2是其他昆虫OBP的同源物，Sgua-CSP1与先前描述的CSP具有高度同源性。
