Cloning and Spatiotemporal Expression Analysis of Bombyx mori elav, an Embryonic Lethal Abnormal Visual Gene

  • Wang, Geng-Xian (National Engineering Laboratory of Modern Silk, Department of Applied Biology, Medical College, Soochow University) ;
  • Liu, Ying (Bioengineering Department, Jiangsu Polytechnic College of Agriculture and Forestry) ;
  • Sim, Yang-Hu (National Engineering Laboratory of Modern Silk, Department of Applied Biology, Medical College, Soochow University) ;
  • Zhang, Sheng-Xiang (College of Forestry, Shandong Agricultural University) ;
  • Xu, Shi-Qing (National Engineering Laboratory of Modern Silk, Department of Applied Biology, Medical College, Soochow University)
  • 발행 : 2009.06.30

초록

Embryonic lethal abnormal visual (elav) is a lethal gene in Drosophila inducing the abnormal development and function of nervous system. We cloned a Bm-elav gene by bioinformatics and biological experiment, based on sequence of ELAV protein and dbEST of Bombyx mori. The full-length of Bm-elav cDNA is 1498 bp, contains a 906 bp open read frame (ORF) encoding a precursor of 301 amino acid residues with a calculated molecular weight of 34 kDa and pI of 8.99. Bm-ELAV protein precursor contains three RNA recognition motifs (RRM) in $24{\sim}91$, $110{\sim}177$ and $222{\sim}295$ bit amino acid residues respectively, and belongs to RNA-binding protein family. Bm-ELAV shared varying positives, ranging from 56% to 60% (Identities from 41% to 45%), with RRM from other species of Xenopus tropicalis, Apis mellifera, Tribolium castaneum, Branchiostoma belcheri and Drosophila. Gene localization indicated that Bm-elav is a single-copy gene, gene mapping within 12-chromosome from 7916.68 knt to 7918.16 knt region of nscaf2993. Spatiotemporal expressions pattern analysis revealed that Bm-elav expressed higher in most tested tissues and developmental stages in whole generation, such as silk gland, fat body, midgut, hemopoietic organ and ovary, but almost no expression in terminated diapause eggs. This suggested that the expression of Bm-elav in early developmental embryonic stages might induce abnormal development like in Drosophila. Cloning of the Bm-elav gene enables us to test its potential role in controlling pests by transferring the gene into field lepidopteran insects in the future.

키워드

참고문헌

  1. Adams MD, Celniker SE, Holt RA, Evans CA, Gocayne JD, Amanatides PG, Scherer SE, Li PW, Hoskins RA, Galle RF, George RA, Lewis SE, Richards S, Ashburner M, Henderson SN, Sutton GG, Wortman JR, Yandell MD, Zhang Q Chen LX, Brandon RC, Rogers YH, Blazej RG, Champe M, Pfeiffer BD, Wan KH, Doyle C, Baxter EG, Helt G, Nelson CR, Gabor GL, Abril JF, Agbayani A, An HJ, Andrews-Pfannkoch C, Baldwin D, Ballew RM, Basu A, Baxendale J, Bayraktaroglu L, Beasley EM, Beeson KY, Benos PV, Berman BP, Bhandari D, Bolshakov S, Borkova D, Botchan MR, Bouck J, Brokstein P, Brottier P, Burtis KC, Busam DA, Butler H, Cadieu E, Center A, Chandra I, Cherry JM, Cawley S, Dahlke C, Davenport LB, Davies P, de Pablos B, Delcher A, Deng Z, Mays AD, Dew I, Dietz SM, Dodson K, Doup LE, Downes M, Dugan-Rocha S, Dunkov BC, Dunn P, Durbin KJ, Evangelista CC, Ferraz C, Ferriera S, Fleischmann W, Fosler C, Gabrielian AE, Garg NS, Gelbart WM Glasser K, Glodek A, Gong F, Gorrell JH, Gu Z, Guan P, Harris M, Harris NL, Harvey D, Heiman TJ, Hernandez JR, Houck J, Hostin D, Houston KA, Howland TJ, Wei MH, Ibegwam C, Jalali M, Kalush F, Karpen GH, Ke Z, Kennison JA, Ketchum KA, Kimmel BE, Kodira CD, Kraft C, Kravitz S, Kulp D, Lai Z, Lasko P, Lei Y, Levitsky AA, Li J, Li Z, Liang Y, Lin X, Liu X, Mattei B, McIntosh TC, McLeod MP, McPherson D, Merkulov G, Milshina NV, Mobarry C, Morris J, Moshrefi A, Mount SM, Moy M, Murphy B, Murphy L, Muzny DM, Nelson DL, Nelson DR, Nelson KA, Nixon K, Nusskern DR, Pacleb JM, Palazzolo M, Pittman GS, Pan S, Pollard J, Puri V, Reese MG, Reinert K, Remington K, Saunders RD, Scheeler F, Shen H, Shue BC, Sidén-Kiamos I, Simpson M, Skupski MP, Smith T, Spier E, Spradling AC, Stapleton M, Strong R, Sun E, Svirskas R, Tector C, Turner R, Venter E, Wang AH, Wang X, Wang ZY, Wassarman DA, Weinstock GM, Weissenbach J, Williams SM, Woodage T, Worley KC, Wu D, Yang S, Yao QA, Ye J, Yeh RF, ZaveriJS, Zhan M, Zhang G, Zhao Q, Zheng L, Zheng XH, Zhong FN, Zhong W, Zhou X, Zhu S, Zhu X, Smith HO, Gibbs RA, Myers EW, Rubin GM, Venter JC (2000) The genome sequence of Drosophila melanogaster. Science 287, 2185-2195 https://doi.org/10.1126/science.287.5461.2185
  2. Antic D, Keene JD (1997) Insights from model systems:Embryonic lethal abnormal visual RNA-binding proteins involved in growth, differentiation, and posttranscriptional gene expression. Am J Hum Genet 61, 273-278 https://doi.org/10.1086/514866
  3. Borgeson CD and Samson ML (2005) Shared RNA-binding sites for interacting members of the Drosophila ELAV family of neuronal proteins. Nucleic Acids Res 33, 6372-6383 https://doi.org/10.1093/nar/gki942
  4. Burd CG, Dreyfuss G (1994) Conserved structures and diversity of functions of RNA-binding proteins. Science 265, 615-621 https://doi.org/10.1126/science.8036511
  5. Campos AR, Rosen DR, Robinow SN, White K (1987) Molecular analysis of the locus elav in Drosophila melanogaster: a gene whose embryonic expression is neural specific. EMBO J 6, 425-431
  6. Gill RW, Sanseau P (2000) Rapid in silico cloning of genes using expressed sequence tags (ESTs). Biotechnol Annu Rev 5, 25-44 https://doi.org/10.1016/S1387-2656(00)05031-6
  7. Goldsmith MR (1995) Molecular Model Systems in the Lepidoptera. Cambridge University Press (ed.), pp. 21-76. Cambridge
  8. Gong P, Epton MJ, Fu GL, Scaife S, Hiscox A, Condon KC, Condon GC, Morrison NI, Kelly DW, Dafa’alla T, Coleman PG, Alphey L (2005) A dominant lethal genetic system for autocidal control of the Mediterranean fruit fly. Nature Biotechnol 23, 453-456 https://doi.org/10.1038/nbt1071
  9. Good PJ (1995) Conserved family of elav-like genes in vertebrates. Proc Natl Acad Sci USA 92, 4557-4561 https://doi.org/10.1073/pnas.92.10.4557
  10. Manley GT, Smitt PS, Dalmau J, Posner JB (1995) Hu antigens:reactivity with Hu antibodies, tumor expression, and major immunogenic sites. Ann Neurol 38, 102-110 https://doi.org/10.1002/ana.410380117
  11. Miao XX, Xub SJ, Li MH, Li MW, Huang JH, Dai FY, Marino SW, Mills DR, Zeng P, Mita K, Jia SH, Zhang Y, Liu WB, Xiang H, Guo QH, Xu AY, Kong XY, Lin HX, Shi YZ, Lu G, Zhang X, Huang W, Yasukochi Y, Sugasaki T, Shimada T, Nagaraju J, Xiang ZH, Wang SY, Goldsmith MR, Lu C, Zhao GP, Huang YP (2005) Simple se-quence repeat-based consensus linkage map of Bombyx mori. Proc Natl Acad Sci USA 102, 16303-16308 https://doi.org/10.1073/pnas.0507794102
  12. Mita K, Kasahara M, Sasaki S, Nagayasu Y, Yamada T, Kanamori H, Namiki N, Kitagawa M, Yamashita H, Yasukochi Y, Kadono-Okuda K, Yamamoto K, Ajimura M, Ravikumar G, Shimomura M, Nagamura Y, Shin-i T, Abe H, Shimada T, Morishita S, Sasaki T (2004) The genome sequence of silkworm, Bombyx mori. DNA Research 11, 27-35 https://doi.org/10.1093/dnares/11.1.27
  13. Robinow S, Campos AR, Yao KM, White K (1988) The elav gene product of Drosophila, required in neurons, has three RNP consensus motifs. Science 242, 1570-1572 https://doi.org/10.1126/science.3144044
  14. Robinow S, White K (1991) Characterization and spatial distribution of the ELAV protein during Drosophila melanogaster development. J Neurobiol 22, 443-461 https://doi.org/10.1002/neu.480220503
  15. Simionato E, Barrios N, Duloquin L, Boissonneau E, Lecorre P, Agnes F (2007) The Drosophila RNA-binding protein ELAV is required for commissural axon midline crossing via control of commissureless mRNA expression in neurons. Dev Biol 301, 166-177 https://doi.org/10.1016/j.ydbio.2006.09.028
  16. Thomas DT, Donnelly CA, Wood RJ, Alphey LS (2000) Insect population control using a dominant, repressible, lethal genetic system. Science 287, 2474-2476 https://doi.org/10.1126/science.287.5462.2474
  17. Xia Q, Zhou Z, Lu C, Cheng D, Dai F, Li B, Zhao P, Zha X, Cheng T, Chai C, Pan G, Xu J, Liu C, Lin Y, Qian J, Hou Y, Wu Z, Li G, Pan M, Li C, Shen Y, Lan X,Yuan L, Li T, Xu H, Yang G, Wan Y, Zhu Y, Yu M, Shen W, Wu D, Xiang Z, Yu J, Wang J, Li R, Shi J, Li H, Li G, Su J, Wang X, Li G, Zhang Z, Wu Q, Li J, Zhang Q, Wei N, Xu J, Sun H, Dong L, Liu D, Zhao S, Zhao X, Meng Q, Lan F, Huang X, Li Y, Fang L, Li C, Li D, Sun Y, Zhang Z, Yang Z, Huang Y, Xi Y, Qi Q, He D, Huang H, Zhang X, Wang Z, Li W, Cao Y, Yu Y, Yu H, Li J, Ye J, Chen H, Zhou Y, Liu B, Wang J, Ye J, Ji H, Li S, Ni P, Zhang J, Zhang Y, Zheng H, Mao B, Wang W, Ye C, Li S, Wang J, Wong GK, Yang H, Biology Analysis Group (2004) A draft sequence for the genome of the domesticated silkworm (Bombyx mori). Science 306, 1937-1940 https://doi.org/10.1126/science.1102210