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Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/30685
標題: cDNA cloning and expression of the vitellogenin receptor from the oriental fruit fly, Bactrocera dorsalis
東方果實蠅之卵黃原蛋白受器cDNA序列分析及表現情形探討
作者: Lin, Wei-Jen
林威仁
關鍵字: Bactrocera dorsalis;東方果實蠅;vitellogenin receptor;ovary;fat body;卵黃原蛋白受器;卵巢;脂肪體
出版社: 昆蟲學系所
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Oocyte membrane localization of vitellogenin receptor coincides with queen flying age, and receptor silencing by RNAi disrupts egg formation in fire ant virgin queens. FEBS J 276: 3110-3123. Mullberg J, Rauch CT, Wolfson MF, Castner B, Fitzner JN, Otten-Evans C, Mohler KM, Cosman D, Black RA. 1997. Further evidence for a common mechanism for shedding of cell surface proteins. FEBS Lett 401: 235–238. Raikhel AS, Dhadialla TS.1992. Accumulation of yolk proteins in insect oocytes. Annu Rev Entomol 37: 217–251. Rajaratnam VS. 1996. Isolation, characterization and complete nucleotide sequence of a Galleria mellonella cDNA coding for the follicle cell-specific yolk protein, YP4. Insect Biochem Molec 26: 545–555. Rodenburg KW, Smolenaars MMW, Hoof DV, Horst DJV-D. 2006. Sequence analysis of the non-recurring C-terminal domains shows that insect lipoprotein receptors constitute a distinct group of LDL receptor family members. Insect Biochem Molec 36: 250-263. 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Schonbaum CP, Perrino JJ, Mahowald AP. 2000. Regulation of the vitellogenin receptor during Drosophila melanogaster Oogenesis. Mol Biol Cell 11: 511–521. Seehuus SC, Norberg K, Gimsa U, Krekling T, Amdam GV. 2006. Reproductive protein protects functionally sterile honey bee workers from oxidative stress. P Natl Acad Sci USA 103: 962–967. Shu YH, Wang JW, Lu K, Zhou JL, Zhou Q, Zhang GR. 2011. The first vitellogenin receptor from a Lepidopteran insect: molecular characterization, expression patterns and RNA interference analysis. Insect Mol Biol 1: 61-73. Snigirevskaya ES, Hays AR, Raikhel AS. 1997. Secretory and internalization pathways of mosquito yolk protein precursors. Cell Tissue Res 290: 129–142. Springer TA. 1998. An extracellular beta-propeller module predicted in lipoprotein and scavenger receptors, tyrosine kinases, epidermal growth factor precursor and extracellular matrix components. J Mol Biol 283: 837–862. Tufail M, Takeda M. 2005. 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摘要: 
卵黃原蛋白透過受器媒介內吞作用吸收到卵內儲存供胚胎發育。本研究選殖東方果實蠅 (Bactrocera dorsalis Hendel) 卵黃原蛋白受器cDNA 序列並分析其表現情形。利用RACE得到一全長6585 bp之cDNA，其開放讀架 (open reading frame, ORF) 可轉譯出1923個胺基酸，其功能區域排列符合昆蟲卵黃原蛋白受器之特性，與黃果蠅 (Drosophila melanogaster) 有極高之相似度 (70%)。利用RT-PCR分析，脂肪體和卵巢兩者皆會表現卵黃原蛋白受器mRNA。進一步以西方墨點法分析，顯示卵巢與脂肪體皆會表現卵黃原蛋白受器之蛋白質 (約213 kDa)。利用qRT-PCR分析於不同日齡 (羽化後十天內) 雌成蟲脂肪體內卵黃原蛋白受器mRNA之表現，結果顯示羽化後至六日齡會隨時間而增加，之後則會隨時間而下降；蛋白質表現則為羽化初期 (羽化後第零天到第二天) 表現量較低，第三天開始增加直到第六天有高量表現，隨後表現量開始下降。qRT-PCR分析於不同日齡 (羽化後十天內) 之雌成蟲卵巢內卵黃原蛋白受器mRNA之表現結果顯示，雌成蟲羽化後有高量表現 (二十四小時內) 之後持續下降直到第二天，第三天開始有上升之趨勢直到第十天。蛋白質的表現則為羽化初期之蛋白表現量較低，從第三天開始有上升的趨勢直到第十天。本研究結果指出卵黃原蛋白受器於卵巢發育時期會持續的表現，而其於脂肪體之功能可能為吸收血體液中之卵黃原蛋白。

The vitellogenin (Vg) is absorbed into the growing oocytes via receptor-mediated endocytosis for the embryonic development. In this study, vitellogenin receptor cDNA of the oriental fruit fly (Bactrocera dorsalis Hendel) was cloned and its expression was analyzed. Using RACE technology, a putative B. dorsalis vitellogenin receptor (BdVgR) was cloned. The BdVgR cDNA has a length of 6585 bp encoding 1923 amino acids. The BdVgR has conserved motif arrangement with other insect VgRs, and showed the high similarity to Drosophila VgR (70%). The PCR analysis showed that the mRNA of BdVgR was expressed in both ovary and fat body. The western blot analysis demonstrated that the BdVgR (~213 kDa) was expressed in both ovary and fat body. The qRT-PCR analysis showed that the expression of BdVgR in the fat body was low after eclosion.The BdVgR mRNA increased from day 4, and it showed the highest level on day 6, and then it decreased with time. The expression level of BdVgR in the ovary was high in 24h after eclosion, and then it decreased until day 2. It increased with time on day 3. The BdVgR protein was expressing highly in the fat body between day3 and day 6; but it gradually increased from day 3 in the ovary. In conclusions, the BdVgR was expressed during the ovary development. The function of BdVgR in the fat body could be involved in reuptake vitellogenins from the hemolymph.
URI: http://hdl.handle.net/11455/30685
其他識別: U0005-1208201102433300
Appears in Collections:昆蟲學系

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