Bioinformatic analysis of Vibrio parahaemolyticus thermolabile hemolysin gene

Abstract

Abstract: Objective To carry out bioinformatic analysis of Vibrio parahaemolyticus thermolabile hemolysin gene(tlh) obtained by PCR amplification.Methods The tlh gene amplified by PCR was cloned into the vector pET32a⁺ and sequenced,followed by analysis of the biological information by with presenting the sequences to the websites of bioinformatics on the Internet.Results and Conclusions The sequenced tlh gene (named tlh14-90) was entered into GenBank with the accession number of AY289609. Tlh14-90 has a length of 1 257 bp with both start and stop codons,having 99% homology with the tlh gene of WP1. Tlh14-90 is predicted to encode a protein containing 418 amino acids (named TLH14-90,with the molecular formula of C₂₁₃₁H₃₁₈₄N₅₄₈O₆₄₉S₁₆,molecular weight of 47 392.9,and the theoretical PI of 4.92). This protein consists of 4 Cys and the contents of Ala,Leu and Asn are 11.0%,7.4% and 7.2%,respectively,having a hydrophobic parameter of -34 calculated using κ-D method. Predicted as a α/□protein for its secondary structure,TLH14-90 has not been identified for its tertiary structure.

CLC Number:

LI Zhi-Feng, NIE Jun, DAI Ying-Chun, LI Jian-dong, CHEN Qing, YU Shou-yi. Bioinformatic analysis of Vibrio parahaemolyticus thermolabile hemolysin gene[J]. Journal of Southern Medical University, 2004, 24(10): 1153-1155.

References

[1] Miyamoto Y,Kato T,Obara Y,et al.In vitro hemolytic characteristic of Vibrio parahaemolyticus:its close correlation with human pathogenicity[J].J Bacteriol,1969,100(2):1147-9.
[2] Okuda JM,Ishibashi E,Hayakawa T,et al.Emergence of a unique O3:K6 clone of Vibrio parahaemolyticus in Calcutta,India,and isolation of strains from the same clonal group from Southeast Asia travelers arriving in Japan[J].J Clin Microbiol,1997,35(12):3150-5.
[3] Takahashi A,Kenjyo N,Imura K,et al.Cl(-) secretion in colonic epithelial cells induced by the Vibrio parahaemolyticus hemolytic toxin related to thermostable direct hemolysin [J].Infect Immun,2000,68(9):5435-8.
[4] Qiao HL,Zhang J.Test and assay the pathogenicity of Vibrio parahaemolyticus [J].Aquat Sci,1999,18(4):29-32.
[5] Taniguchi H,kubomura S,Hirano H,et al.Cloning and characterization of gene encoding a new thermostable hemolysin from Vibrio parahaemolyticus [J].FEMS Microbiol Lett,1990,55 (3):339-45.
[6] Yanagase Y,Inoue K,Ozaki M,et al.Hemolysins and related enzymes of Vibrio parahaemolyticus.I.Identification and partial purification of enzymes[J].Biken J,1970,13(2):77-92.
[7] 李志峰,聂军.副溶血弧菌tlh基因克隆载体和表达载体的构建[J].中国公共卫生,2003,19(4):418-9.Li ZF,Nie J.Construction of clone and expression vector of gene of Vibrio parahaemolyticus [J].Chin J Pub Health,2003,19(4):418-9.
[8] 苏建新,聂军,吴振龙.副溶血性弧菌耐热直接相关溶血素基因的克隆与序列分析[J].第一军医大学学报,2002,22(6):515-7.Su JX,Nie J,Wu ZL.Cloning and sequence analysis of thermostable direct hemolysin-related hemolysin gene of Vibrio parahaemolyticus[J].J First Mil Med Univ/Di Yi Jun Yi Da Xue Xue Bao,2002,22(6):515-7.
[9] 欧阳曙光,贺福初.生物信息学:生物实验数据和计算技术结合的新领域[J].科学通报(Sci Bull),1999,44(14):1457-68.
[10] Shinoda S,Matsuoka H,Tsuchie T,et al.Purification and characterizaton of a lecithin-haemolysin from Escherichia coli transformed by a Vibrio parahaemolyticus gene[J].J Gen Microobiol,1991,137(Pt 12):2705-11.
[11] 李明,谢毅,李英杰,等.恶性疟原虫海南、云南、安徽株裂殖子表面抗原MSA2基因的克隆和序列分析[J].第一军医大学学报,1995,15(2):94-8.Li M,Xie Y,Li Y J,et al.Cloning and sequence analysis of the gene encoding the second region of MSA1 from three Chinese isolates[J].J First Mil Med Univ/Di Yi Jun Yi Da Xue Xue Bao,1995,15(2):94-8.
[12] Ellison RK,Malnati E,Depaloa A,et al.Population of vibrio parahoemolyticus in retail oysters from Florida using two metods[J].J Food Prot,2001,64(5):682-6.
[13] Myers ML,Panicker G,Bej AK.PCR detection of a newly emerged pandemic Vibrio parahaemolyticus O3:K6 pathogen in pure cultures and seeded waters from the gulf of Mexico [J].Appl Environ Microbiol,2003,69(4):2194-200.
[14] Iida T,Park KS,Suthienkul O,et al.Close proximity of tdh,trh and ure genes on the chromosone of Vibrio parahaemolyticus [J].Microbiology,1998,144(pt 9):2517-23.
[15] Takahasi A,Kenjyo N,Imura K,et al.Secretion in colonic epithelial cell induced by the vibrio parahaemalyticus hemolytic toxin related to thermostable direct hemolysin [J].Infec Immun,2000,68(9):5435-8.