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研究生:陳秀玲
研究生(外文):Hsiu-Lin Chen
論文名稱:東方果實蠅卵黃原蛋白基因之選殖與其構造及表現分析
論文名稱(外文):Cloning and structural and expression analysis of vitellogenin genes of the oriental fruit fly, Bactrocera dorsalis (Hendel)
指導教授:路光暉
指導教授(外文):Kuang-Hui Lu
學位類別:碩士
校院名稱:國立中興大學
系所名稱:昆蟲學系
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:87
中文關鍵詞:卵黃原蛋白基因表現東方果實蠅選殖
外文關鍵詞:vitellogeningene expressionBactrocera dorsaliscloning
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以第六日齡東方果實蠅 (Bactrocera dorsalis (Hendel)) 雌成蟲腹部總量RNA進行反轉錄作用 (reverse transcription) 和聚合酵素連鎖反應 (polymerase chain reaction, PCR),得到兩卵黃原蛋白基因序列,稱為Bdvg1與Bdvg2。Bdvg1為具1507 bp之cDNA,而Bdvg2 具1434 bp,兩者皆具完整之開放讀架(opening reading frame)。將Bdvg1與Bdvg2分別與親緣關係相近之雙翅目昆蟲如地中海果實蠅 (Ceratitis capitata)、加勒比海果實蠅(Anastrepha suspensa)、黃果蠅(Drosophila melanogaster)、綠頭蒼蠅 (Calliphora erythrocephala)與家蠅(Musca domestica)之卵黃原蛋白基因序列比對,發現Bdvg1與Bdvg2與上述昆蟲之卵黃原蛋白基因序列之相似度、一致性皆有60%以上程度相似,因此更確認所得之Bdvg1與Bdvg2應為東方果實蠅之卵黃原蛋白基因。再者Bdvg1與Bdvg2的相似度與一致性更高達71.0和70.9﹪,同時比對東方果實蠅的卵黃原蛋白基因Bdvg1與Bdvg2之胺基酸序列,亦顯示此兩基因之胺基酸序列非常相似,將之與其他雙翅目環列亞目昆蟲之卵黃原蛋白比較,結果顯示這些卵黃原蛋白胺基酸序列,靠近C端的部分具GLARGDADFDAIHTSTCGMGT、PGNERNFPAV及GKRAYMGIATD等序列保留區域(conversed region),N端則是各卵黃原蛋白間變異較大的區域。目前,於卵黃原蛋白基因體DNA構造分析上,已篩選到一個長約1.5 kb的菌株並已定序,發現該片段與卵黃蛋白基因Bdvg 1有約708 bp的重疊區域。在此片段中存在一個長度為67 bp的intron,且intron之5’端與3’端分別保有GT與AG之保留序列;另外在-27至-22的位置具有TATAAA序列,此為TATA box保留序列,而在+3至+7的位置 處具一ACAGT序列,此為capping site 保留序列。
東方果實蠅的卵黃原蛋白基因只於雌蟲的成蟲期表現,於卵、幼蟲、蛹等時期則無表現之趨勢。另外可發現Bdvg1和Bdvg2兩基因在卵巢細胞中的表現量接遠低於頭、胸部和脂肪體細胞。同時,在雄蟲中並無偵測出卵黃蛋白基因之表現。就脂肪體細胞而言,在第一日齡的雌成蟲即可探測出Bdvg1之微量表現,而隨著成蟲日齡增加該基因有逐漸增加表現的趨勢;Bdvg2則是在第二日齡開始有微量表現,而兩基因到了第五日齡皆可清楚看出其表現,並有隨即表現減弱的傾向。而在偵測卵巢細胞中的卵黃蛋白基因表現上,Bdvg1從第二日齡時可以開始偵測出其表現並且逐漸明顯表現,但是到第六日齡時卻大大減弱其表現量,之後再隨著日齡增加逐漸有表現量加強現象。相同的現象出在Bdvg2於卵巢細胞中的表現情形。

In this study, two vitellogenin genes from oriental fruit fly, Bactrocera dorsalis, named Bdvg1 and Bdvg2, respectively, were identified. Bdvg1 was 1507 bp long and contained an open reading frame encoding 437 amino acids. The identities of Bdvg1 were 78.7﹪to CcVG1 of Ceratitis capitata, 76.9﹪to AsYP of Anastrepha suspensa, 67.6﹪to CeYP3 of Calliphora erythrocephala, 65.0﹪to CcVG2 of C. capitata and CeYP2 of C. erythrocephala, respectively. Bdvg2 was 1434 bp long and contained an open reading frame as well encoding 440 amino acids. The identities of Bdvg2 were 71.9﹪to CcVG1 of C. capitata, 70.5﹪to AsYP of A. suspensa, 65.0﹪to CeYP3 of C. erythrocephala, 64.8﹪to CcVG2 of C. capitata and 64.5﹪to CeYP2 of C. erythrocephala. Bdvg1 and Bdvg2 were highly homologous to the other dipteran vitellogenin genes. The carboxyl-end domains of Bdvg1 and Bdvg2 were much conserved in comparison with other fruit flies. However, the amino-end domains were much diversed. The signal sequences of Bdvg1 and Bdvg2 are similar to the signal sequences of vitellogenins of D. melanogaster, C. capitata and C. erythrocephala.
1.5 kb genomic DNA fragment of Bdvg1 was cloned and sequenced. The sequence of this fragment overlapped with the 5’-end of the Bdvg1. The fragment contains an intron that is 67 bp downstream of Bdvg1, and a TATA box and a cap site conserved sequences were found at locatins of —27 to —22 and +3 to +7, respectively.
Rt-PCR studies revealed that the vitellogenin genes in B. dorsalis expressed specifically in the adult females but not in males, eggs, larvae and pupae. The Bdvg1 and Bdvg2 expression in ovaries of female adult B. dorsalis significantly increased since day 2 and after. And the Bdvg1 expression in fat body of female adult B. dorsalis significantly increased since day 1 and after but day 2 in Bdvg2. Furthermore, Bdvg1 and Bdvg2 transcripts were detected in the heads, thoraxes and abdomens of 6-day-old adult females.

中文摘要 ----------------------------------------------1
英文摘要 ----------------------------------------------3
前言 --------------------------------------------------5
前人研究 ----------------------------------------------7
材料與方法 -------------------------------------------18
結果 -------------------------------------------------30
討論 -------------------------------------------------37
參考文獻 ---------------------------------------------47
圖 ---------------------------------------------------54
表 ---------------------------------------------------80
附錄 -------------------------------------------------82

Abrahamsen, N., A. Martinez, T. Kjar, L. Sondergaard and M. Bownes 1993 Cis-regulatory sequences leading female-specific expression of yolk protein genes 1 and 2 in the fat body of Drosophila melanogaster. Mol. Gen. Genet. 237: 41-48.
Adams, T. S. and P. A. Filipi 1985 Vitellin and vitellogenin concentrations during oogenesis in the house fly, Musca domestica. J. Insect Physiol. 29: 723-733.
Agui, N., S. Izumi and S. Tomino 1985 The role of ecdysteroids and juvenoids in vitellogenin levels and follicle development in the housefly, Musca domestica. Appl. Entomol. Zool. 20: 179-188.
Agui, N., T. Shimada, S. Izumi and S. Tomino 1991 Hormonal control of vitellogenin mRNA levels in the male and female housefly, Musca domestica. J. Insect Physiol. 37: 383-390.
Barnett, T., C. Pachl, J. P. Gergen and P. C. Wensink 1980 The isolation and characterization of Drosophila yolk protein genes. Cell 21: 729-738.
Belote, J. M., A. M. Handler, M. F. Wolfner, K. J. Livak and B. Baker 1985Sex-specific regulation of yolk protein gene expression in Drosophila. Cell 40: 339-348.
Bownes, M. 1982 Hormonal and genetic regulation of vitellogenesis in Drosophila. Q. Rev. Biol. 57: 247-274.
Bownes, M. 1992 Why is there sequence similarity between insect yolk proteins and vertebrate lipases? J. Lipid Res. 33: 777-789.
Bownes, M. 1994 The regulation of the yolk protein genes, a family of sex differentiation genes in Drosophila melanogaster. BioEssays 16: 745-752.
Bownes, M. and B. D. Hames 1978 Analysis of the yolk protein in Drosophila melanogaster. Translation in a cell free system and peptide analysis. FEBS Lett. 96: 327-330.
Bownes, M., E. Ronaldson, D. Mauchline and A. Martinez 1993Regulation of vitellogenesis in Drosophila. Int. J. Insect Morphol. Embryol. 22: 349-367.
Brennan, M. D., A. J. Weiner, T. J. Goralski and A. P. Mahowald 1982 The follicle cells are a major site of vitellogenin synthesis in Drosophila melanogaster. J. Dev. Biol. 89: 225-236.
Burtis, K. C., K. T. Coschigano, B. S. Baker and P. C. Wensink 1991The doublesex proteins of Drosophila melanogaster bind directly to a sex-specific yolk protein gene enhancer. J. EMBO 10: 2577-2582.
Chen, J. S. and A. S. Raikhel 1996 Subunit cleavage of mosquito pro-vitellogenin by a subtilisin-like convertase. Proc. Natl. Acad. Sci. USA 93: 6186-6190.
Chen, J. S., W. L. Cho and A. S. Raikhel 1994 Analysis of mosquito vitellogenin cDNA similarity with vertebrate phosvitins and arthropod serum regions. J. Mol. Biol. 237: 641-647.
Chen, J. S., T. W. Sappington and A. S. Raikhel 1997 Extensive sequence conservation among insect, nematode, and vertebrate vitellgenins reveals ancient common ancestry. J. Molec. Evol. 44: 440-451.
Chiu, H. T. 1978 Studies on the improvement of mass rearing for oriental fruit flies. Plant Protect. Bullet. 20: 87-92 (in Chinese).
Comas, D., M. D. Piulachs and X. Belles 1999 Fast induction of vitellogenin gene expression by juvenile hormone III in the cockroach Blattella germanica (L.) (Dictyoptera, Blattellidae). Insect Biochem. Molec. Biol. 29: 821-827.
Comas, D., M. D. Piulachs and X. Belles 2000 Vitellogenin of Blattella germanica (L.) (Dictyoptera, Blattellidae) : nucleotide sequence of the cDNA and analysis of the protein primary structure. Arch. Insect Biochem. Physiol. 45: 1-11.
De Bianchi, A. G., M. Coutiho, S. D. Pereira, O. Marinotti and H. J. Targa 1985 Vitellogenin and vitellin of Musca domestica quantification and synthesis by fat bodies and ovaries. Insect Biochem. 15: 77-84.
Dhadialla, T. S. and A. S. Raikhel 1990 Biosynthesis of mosquito vitellogenin. J. Biol. Chem. 265: 9924-9933.
Dhadialla, T. S., A. R. Hays and A. S. Raikhel 1992 Characterization of the solubilized mosquito vitellogenin receptor. Insect Biochem. Molec. Biol. 22: 803-816.
Djordjevic, J. T., S. Bieri, R. Smith and P. A. Kroon 1996 A deletion in the first cysteine-rich repeat of the low-density-lipoprotein receptor leads to the formation of multiple misfolded isomers. Eur. J. Biochem. 239: 214-219.
DonWheeler, G. and F. Engelmann 1997 The biosynthesis and processing of vitellogenin in the fat bodies of females and males of the cockroach Leucophaea maderae. Insect Biochem. Molec. Biol. 27: 901-918.
Edwards, G. C., R. P. Braun and G. R. Wyatt 1993 Induction of vitellogenin synthesis in Locusta migratoria by the juvenile hormone analog, pyriproxyfen. J. Insect Physiol. 39: 609-614.
Garabedian, M. J., A. D. Shirras, M. Bownes and P. C. Wensink 1987 The nucleotide of the gene coding for Drosophila melanogaster yolk protein 3. Gene 55: 1-8.
Giorgi, F., J. T. Bradley and J. H. Nordin 1999 Differential vitellin polypeptide processing in insect embryos. Micron 30: 579-596.
Glinka, A. V. and G. R. Wyatt 1996 Juvenile hormone activation of gene transcription in locust fat body. Insect Biochem. Molec. Biol. 26: 13-18.
Grundemann, D. and H. Koepsell 1994 Ethidium bromide staining during denaturation with glyoxal for sensitive detection of RNA in agarose gel electrophoresis. Anal. Biochem. 216: 459-461.
Hander, A. M. 1997Developmental regulation of yolk protein gene expression in Anastrepha suspensa. Arch. Insect Biochem. Physiol. 36: 25-35.
Handler, A. M. and P. D. Shirk 1988Identification and analysis of the major yolk polypeptide from the caribbean fruit fly, Anastrepha suspensa (Loew). Arch. Insect Biochem. Physiol. 9: 91-106.
Harnish, D. G. and B. N. White 1982a Insect vitellins: identification, purification, and characterization from eight orders. J. Exp. zool. 220: 1-10.
Harnish, D. G. and B. N. White 1982b An evolutionary model for the insect vitellins. J. Mol. Evol. 18: 405-413.
Harshman, L. G. 1998 Differential gene expression in insect: transcription control. Annu. Rev. Entomol. 43: 671-700.
Heilmann, L. J., P. M. Trewitt and K. Kumaran 1993 Proteolytic processing of the vitellogenin precursor in the boll weevil, Anthonomus grandis. Arch. Insect Biochem. Physiol. 23: 125-134.
Hirai, M., D. Watanabe, A. Kiyota and Y. Chinzei 1998 Nucleotide sequence of vitellogenin mRNA in the bean bug, Riptorus clavatus: analysis of processing in the fat body and ovary. Insect Biochem. Molec. Biol. 28: 537-547.
Houseman, J. G. and P. E. Morrison 1986 Absence of female-specific protein in the hemolymph of stable fly Stomoxys calcitrans (L.) (diptera: muscidae). Arch. Insect Biochem. Physiol. 3: 205-213.
Hovemann, B., R. Galler, U. Walldorf, H. Jupper and E. K. F. Bautz 1981 Vitellogenin in Drosophila melanogaster: sequence of the yolk protein 1 gene and its flanking regions. Nucleic Acids Res. 9: 4721-4734.
Hung, M. C. and P. C. Wensink 1981 The sequence of the Drosophila melanogaster gene for yolk protein 1. Nucleic Acids Res. 9: 6407-6419.
Izumi, S., K. Yano, Y. Yamamoto and S. Y. Takahashi 1994 Yolk protein from insect eggs: structure, biosynthesis and programmed degradation during embryogenesis. J. Insect Physiol. 40: 735-746.
Kageyama, Y., T. Kinoshita, Y. Umesono, M. Hatakeyama and K. Oishi 1994 Cloning of cDNA for vitellogenin of Athalia rosae ( Hymenoptera ) and characterization of the vitellogenin gene expression. Insect Biochem. Molec. Biol. 24: 599-605.
Kelly, T. J., T. S. Adams, M. B. Schwartz, M. J. Birnbaum and E. C. Rubemstein 1987 Juvenile hormone and ovarian maturation in the Diptera: a review of recent results. J. Insect Biochem. 17: 1089-1093.
Kjer, K. M., G. D. Baldridge and A. M. Fallon 1994 Mosquito large subunit ribosomal RNA: simultaneous alignment of primary and secondary structure. Bioch. Biophy. Acta. 1217:141-146.
Kunkel, J. G. and J. H. Nordin 1985 Yolk protein. pp83-111. In: Kerbut, G. A. and L. I. Gilbert, eds. Comprehensive Insect Physiology, Biochemistry and Pharmacology. Pergamon Press, Oxford.
Lee, J. M., M. Hatakeyama and K. Oishi 2000a A simple and rapid method for cloning insect vitellogenin cDNAs. Insect Biochem. Molec. Biol. 30: 189-194.
Lee, J. M., N. Y. M. Hatakeyama, T. W. Bae and K. Oishi 2000b Vitellogenin of the cicada Graptopsaltria nigrofuscata (Homoptera): analysis of its primary structure. Insect Biochem. Molec. Biol. 30: 1-7.
Levedakou, E. N. and C. E. Sekeris 1987 Isolation and characterization of vitellin from the fruitfly, Dacus oleae. Arch. Insect Biochem. Physiol. 4: 297-311.
Lin, C. P. 1999 Isolation and identification of a yolk protein gene of the oriental fruit fly, Bactrocera dorsalis (Hendel), Master’s Thesis, Department of Entomlogy, National Chung Hsing University. 68pp (in Chinese).
Loukeris, T. G., I. Livadasas, B. Arca, S. Zabalou and C. Savakis 1995Gene transfer into the medfly, Ceratitis capitata, with a Drosophila hydei transposable element. Science 270: 2002-2005.
Lu, S. L. 2000 Purification of yolk proteins of the oriental fruit fly, Bactrocera dorsalis (Hendel), Bachelor’s Thesis, Department of Entomology, National Chung Hsing University. 22pp (in Chinese).
Martin, D., M. D. Piulachs and X. Belles 1996 Production and extraovarian processing of vitellogenin in ovariectomized Blattella germanica (L.) (Dictyoptera, Blattellidae). J. Insect Physiol. 42: 101-105.
Martin, D., M. D. Piulachs, D. Comas and X. Belles 1998 Isolation and sequence of a partial vitellogenin cDNA from the cockroach, Blattella germanica (L.) (Dictyoptera, Blattellidae), and characterization of the vitellogenin gene expression. Arch. Insect Biochem. Physiol. 38: 137-146.
Martinez, A. and M. Bownes 1994 The sequence and expression pattern of the Calliphora erythrocephala yolk protein A and B genes. J. Mol. Evol. 38: 336-351.
Mayadas, T. N. and D. D. Wagner 1992 Vicinal cysteines in the prosequence play a role in von Willebrand factor multimer assembly. Proc. Natl. Acad. Sci. USA 89: 3531-3535.
Miller, M. S., M. Benore-Parsons and H. B. White 3rd 1982 Dephosphorylation of chicken riboflavin-binding protein and phosvitin decreases their uptake by oocytes. J. Biol. Chem. 257: 6818-6824.
Munroe, D. J., R. Loebbert, E. Bric, T. Whitton and D. Prawitt 1995 Systematic screening of an arrayed cDNA library by PCR. Proc. Natl. Acad. Sci. USA 92: 2209-2213.
Nose, Y., J. M. Lee, T. Ueno, M. Hatakeyama and K. Oishi 1997 Cloning of cDNA for vitellogenin of the parasitoid wasp, Pompla nipponica (Hymenoptera: apocrita: ichneumonidae ) : vitellogenin primary structure and evolutionary considerations. Insect Biochem. Molec. Biol. 27: 1047-1056.
O'Connor, J. L. and M. G. Wade 1991 Control of buffer pH during agaroes gel electrophoresis of glyoxylated RNA. BioTechniques 10: 300-302.
Rina, M. D. and A. C. Mintzas 1988 Biosynthesis and regulation of two vitellogenins in the fat body and ovaries of Ceratitia capitata (Diptera). Roux's Arch. Dev. Biol. 197: 167-174.
Rina, M. and C. Savalis 1991 A cluster of vitellogenin gene in the Mediterranean fruit fly Ceratitis capitata: sequence and structural conservation in Dipteran yolk protein and their genes. Genetics 127: 769-780.
Romans, P., Z. Tu, Z. Ke and H. H. Hagedorn 1995 Anaylsis of a vitellogenin gene of the moquito, Aedes aegypti and comparisons to vitellogenins from other organims. Insect Biochem. Molec. Biol. 25: 939-958.
Sambrook, J. and E. F. Fritsch 2001 Molecular Cloning, A Labortory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New Yolk.
Sappington, T. W. and A. S. Raikhel 1998 Molecular characteristics of insect vitellogenins and vitellogenin receptor. Insect Biochem. Molec. Biol. 28: 277-300.
Sappington, T. W., A. R. Hays and A. S. Raikhel 1995Mosquito vitellogenin receptor: purification, developmental and biochemical characterization. Insect Biochem. Molec. Biol. 25: 807-817.
Shirk, P. D. and O. P. Perera 1998 5' coding region of the follicular epithelium yolk polypeptide 2 cDNA in the moth, Plodia interpunctella, contains an extended coding region. Arch. Insect Biochem. Physiol. 39: 98-108.
Sorge, D., R. Nauen, S. Range and K. H. Hoffmann 2000 Regulation of vitellogenesis in the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae). J. Insect Physiol. 46: 969-976.
Thomas, D. D., C. A. Donnelly, R. J. Wood and L. S. Alphey 2000Insect population control using a dominant, repressible, lethal genetic system. Science 287: 2474-2476.
Tsai, Y. T. 1997 Effects of physiological factors on vitellogenesis and ovarian development in the oriental fruit fly, Bactrocera dorsalis (Hendel). Department of Entomology, National Chung Hsing University. Mastter’s Thesis. 83pp (in Chinese).
Tufail, M., J. M. Lee, M. Hatakeyama, K. Oishi and M. Takeda 2000 Cloning of vitellogenin cDNA of the American cockroach, Periplaneta americana (Dictyoptera), and its structural and expression analysis. Arch. Insect Biochem. Physiol. 45: 37-46.
Wyatt, G. R. 1988 Vitellogenin synthesis and analysis of juvenile hormone action in locust fat body. Can. J. Zool. 66: 2600-2610.
Wyatt, G. R., R. P. Braun and J. Zhang 1996 Priming effect in gene activation by juvenile hormone in locust fat body. Arch. Insect Biochem. Physiol. 32: 633-640.
Yan, Y. L., C. J. Kunert and J. H. Postlethwait 1987 Sequence homologies among the three yolk polypeptide (Yp) genes in Drosophila melanogaster. Nucleic Acids Res. 15: 67-85.
Zhang, J., D. S. Saleh and G. R. Wyatt 1996 Juvenile hormone regulation of an insect gene: a specific transcription factor and a DNA response element. Mol. Cell. Endocrinol. 122: 15-20.
Zongza, V. and G. J. Dimitriadis 1988 Vitellogenesis in the insect Dacus oleae. isolation and characterization of yolk protein mRNA. Insect Biochem. 18: 651-660

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