Cloning and Sequence Analysis of MyoD Gene in Red Tilmpa Oreochromis sp.(O.mossambicus×O.niloticus)

Abstract

Abstract: As a member of myogenic regulatory factors（MRFs）, MyoD play a positive regulation in the process of muscle proliferating.In this study, MyoD cDNA of Red Tilmpa Oreochromis sp.（O.mossambicus×O.niloticus）was obtained using homology cloning strategy and rapid cDNA end amploification. The results showed that the open reading frame（ORF）of MyoD was 903 bp length and encoded 300 amino acids, including basic helix-loop-helix（bHLH）domain which composed of 1^st to 114^th amino acids and 109^th to 165^th amino acids, CDK4 combined structure（199^th to 213^th）and Helix III structure（233^th to 249^th）. The signal peptide was not detected in MyoD. The secondary structure of MyoD protein is HLH dimmer. The phylogenetic trees was constructed based on some fish MyoD ORF and suggested that Oreochromis sp. O. aureaus and O. niloticus were clustered together firstly.

Key words: MyoD , Sequence analysis , Cloning , Red Tilmpa（Oreochromis sp.）

Guo Yihui ,Fan Sigang, Huang Guiju, Zhu Kecheng, Yu Dahui. Cloning and Sequence Analysis of MyoD Gene in Red Tilmpa Oreochromis sp.(O.mossambicus×O.niloticus)[J]. Biotechnology Bulletin, 2014, 0(11): 147-151.

References

[1] Rawls A, Valdez MR, Zhang W, et al. Overlapping functions of the myogenic bHLH genes MRF4 and MyoD revealed in double mutant mice[J]. Development, 1998, 125（13）：2349-2358.
[2] Lassar AB, Buskin JN, Lockshon D, et al. MyoD is a sequence-spe-cific DNA binding protein requiring a region of myc homology to bind to the muscle creatine kinase enhancer[J]. Cell, 1989, 58（5）：823-831.
[3] Blackwell TK, Weintraub H. Differences and similarities in DNA-binding preferences of MyoD and E2A protein complexes revealed by binding site selection[J]. Science, 1990, 250（4984）：1104-1110.
[4] Te Pas MF, Verburg FJ, Gerritsen CL, et al. Messenger ribonucleic acid expression of the MyoD gene family in muscle tissue at slaughter in relation to selection for porcine growth rate[J]. Journal of Animal Science, 2000, 78（1）：69-77.
[5] Lee H, Habas R, Abate-Shen C. MSX1 cooperates with histone H1b for inhibition of transcription and myogenesis[J]. Science, 2004, 304（5677）：1675-1678.
[6] Davis RL, Weintraub H, Lassar AB. Expression of a single transfected cDNA converts fibroblasts to myoblasts[J]. Cell, 1987, 51：987-1000.
[7] Kobiyama A, Nihei Y, Hirayama Y, et al. Molecular cloning and developmental expression patterns of the MyoD and MEF2 families of muscle transcription factors in the carp[J]. Journal of Experimental Biology, 1998, 201（20）：2801-2813.
[8] Tan X, Du SJ. Differential expression of two MyoD genes in fast and slow muscles of gilthead seabream（Sparus aurata）[J]. Dev Genes Evol, 2002, 212（8）：207-217.
[9] 于凌云, 白俊杰, 叶星, 等. 大口黑鲈 MyoD cDNA 的克隆和序列分析[J]. 生物技术通报, 2008（S1）：301-306.
[10] Wang L, Bai J, Luo J, et al. Molecular cloning and expression of grass carp MyoD in yeast Pichia pastoris[J]. Journal of Bioche-mistry and Molecular Biology, 2007, 40（1）：22-28.
[11] Chen YH, Liang CT, Tsai HJ. Expression, purification and DNA-binding activity of tilapia muscle specific transcription factor, MyoD, produced in Escherichia coli[J]. Comparative Biochemistry and Physiology Part B：Biochemistry and Molecular Biology, 2002, 131（4）：795-805.
[12] 卢中华, 俞菊华, 李红霞, 等. 奥利亚罗非鱼, 尼罗罗非鱼 MyoD1 和 MyoD2 基因特征及差异[J]. 中国水产科学, 2010, 17（5）：903-912.
[13] Hall TE, Cole NJ, Johnston IA. Temperature and the expression of seven muscle-specific protein genes during embryogenesis in the Atlantic cod Gadus morhua L[J]. J Exp Biol, 2003, 206（18）：3187-3200.
[14] Petersen TN, Brunak S, von Heijne G, et al. SignalP 4.0：discriminating signal peptides from transmembrane regions[J]. Nature Methods, 2011, 8（10）：785-786.
[15] Kiefer F, Arnolol K, Künzli M, et al. The SWISS-MODEL Reposi-tory and associated resources[J]. Nucleic Acids Research, 2009, 37（Suppl.1）：D387-D392.
[16] Tamura K, Dudley J, Nei M & Kumar S. MEGA4：molecular evolutionary genetics analysis（MEGA）software version 4.0[J]. Molecular Biology and Evolution, 2007, 24（8）：1596-1599.
[17] Soleimani VD, Yin H, Jahani-Asl A, et al. Snail regulates MyoD binding-site occupancy to direct enhancer switching and differentiation-specific transcription in myogenesis[J]. Molecular Cell, 2012, 47（3）：457-468.
[18] 郭奕惠, 喻达辉, 黄桂菊, 等. 中国主要养殖罗非鱼亲缘关系的COI序列分析[J]. 华中农业大学学报, 2009（1）：75-79.
[19] 郭奕惠, 喻达辉, 黄桂菊, 等. 细胞核rDNA序列分析5种罗非鱼的亲缘关系[J]. 安徽农业科学, 2009, 37（8）：3444-3447.