全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

日本鳗鲡I型Cathelicidin基因的克隆与原核表达

DOI: 10.13560/j.cnki.biotech.bull.1985.2015.07.018, PP. 124-131

Keywords: 日本鳗鲡,Cathelicidin,抗菌肽,RACE-PCR

Full-Text   Cite this paper   Add to My Lib

Abstract:

Cathelicidin是目前发现的一个最大抗菌肽家族,具有多重生物学功能。旨在探索和利用鱼类Cathelicidin抗菌机制和潜在的生物学功能。应用RACE-PCR技术获得日本鳗鲡I型Cathelicidin(AjCathI)基因的cDNA全长序列。AjCathI的cDNA全长为842bp,开放阅读框为570bp,编码189个氨基酸。氨基酸序列分析表明,AjCathI靠近C端有4个保守的半胱氨酸序列,抗菌成熟肽与香鱼(Plecoglossusaltivelis)抗菌成熟肽相似性最高,同源性为65.57%。进化分析表明,AjCathI与其它鱼类亲缘关系较近,处于同一个分支上。将AjCathI基因克隆至pET-28a载体,转化EscherichiacoliBL21(DE3)宿主菌进行表达。结果表明,AjCathI以包涵体形式表达,经鎳柱纯化、Westernblot验证和透析复性,获得高纯度的重组蛋白。

References

[1]  Uzzell T, Stolzenberg ED, Shinnar AE, et al. Hagfish intestinal antimicrobial peptides are ancient cathelicidin[J]. Peptides, 2003, 24(11):1655-1667.
[2]  Chang CI, Zhang YA, Zou J, et al. Two cathelicidin genes are present in both rainbow trout(Oncorhynchus mykiss)and Atlantic salmon(Salmo salar)[J]. Antimicrob Agents, 2006, 50(1):185-195.
[3]  Maier VH, Dorn KV, Gudmundsdottir BK, et al. Characterisation of cathelicidin gene family members in divergent fish species[J]. Mol Immunol, 2008, 45(14):3723-3730.
[4]  Broekman DC, Frei DM, Gylfason GA, et al. Cod cathelicidin:Isolation of the mature peptide, cleavage site characterization and developmental expression[J]. Dev Comp Immunol, 2011, 35(3):296-303.
[5]  Scocchi M, Pallavicini A, Salgaro R, et al. The salmonid cathelicidin:A gene family with highly varied C-terminal antimicrobial domains[J]. Comp Biochem Physiol B, 2009, 152(4):376-381.
[6]  Lu XJ, Chen J, Huang ZA, et al. Identification and characterization of a novel cathelicidin from ayu, Plecoglossus altivelis[J]. Fish Shell Immunol, 2011:31(1):52-57.
[7]  Arafa SZ, Reda ES. Surface features of the monogenean gill parasites Pseudodactylogyrus anguillae and Pseudodactylogyrus bini from the European eel Anguilla anguilla in Egypt[J]. Parasitol Res, 2012, 110:127-133.
[8]  Cao HP, He S, Wei RP, et al. Bacillus amyloliquefaciensG1:A potential antagonistic bacterium against eel-pathogenic Aeromonas hydrophila[J]. Evid Based Complement Alternat Med, 2011, 82104:1-7.
[9]  Bergwerff AA, Scherpenisse P. Determination of residues of malachite green in aquatic animals[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2003, 788:351-359.
[10]  Végh AG, Nagy K, Bálint Z, et al. Effect of antimicrobial peptide- amide:indolicidin on biological membranes[J]. J Biomed Biotechnol, 2011:670589.
[11]  Baumann A, Démoulins T, Python S, et al. Porcine cathelicidins efficiently complex and deliver nucleic acids to plasmacytoid dendritic cells and can thereby mediate bacteria-induced IFN-α responses[J]. J Immunol, 2014, 193(1):364-371.
[12]  Garnier J, Gibrat JF, Robson B. GOR method for predicting protein secondary structure from amino acid sequence[J]. Methods Enzymol, 1996, 266:540-553.
[13]  Ramos C, Pietri R, Lorenzo W, et al. Recombinant hemoglobin II from Lucina pectinata:A large-scale method For hemeprotein expression in E. coli[J]. Protein J, 2010, 29(2):143-151.
[14]  Villaverde A, Carril M. Protein aggregation in recombinant bacteria:biological role of inclusion bodies[J]. Biotechnol Lett, 2003, 25(17):1385-1395.
[15]  Ventura S. Sequence determinants of protein aggregation:tools to increase protein solubility[J]. Microb Cell Fact, 2005, 4(1):11-18.
[16]  Zhang JJ, Yan QP, Ji RX, et al. Isolation and characterization of a hepcidin peptide from the head kidney of large yellow croaker, Pseudosciaena crocea[J]. Fish Shell Immunol, 2009, 26:864-870.
[17]  Moon JY, Henzler-Wildman KA, Ramamoorthy A. Expression and purification of a recombinant LL-37 from Escherichia coli[J]. Biochim Biophys Acta, 2006, 1758(9):1351-1358.
[18]  Chen K, Xiang Y, Huang J, et al. The formylpeptide receptor 2(Fpr2)and its endogenous ligand cathelin-related antimicrobial peptide(CRAMP)promote dendritic cell maturation[J]. J BiolChem, 2014, 289(25):17553-17563.
[19]  Lee JY, Yang ST, Kim HJ, et al. Different modes of antibiotic action of homodimeric and monomeric bactenecin, a cathelicidin-derived antibacterial peptide[J]. BMB Rep, 2009, 42(9):586-592.
[20]  Ramanathan B, Davis EG, Ross CR, et al. Cathelicidin:microbicidal activity, mechanisms of action, and roles in innate immunity[J]. Microbes Infect, 2002, 4(3):361-372.
[21]  Geourjon C, Déleage G. SOPMA: Significant improvements in protein secondary structure prediction by consensus prediction from multiple alignments[J]. CABIOS, 1995, 11(6):681-684.
[22]  Frishman D, Argos P. Incorporation of non-local interactions in protein secondary structure prediction from the amino acid sequence[J]. Protein Eng, 1996, 9(2):133-142.
[23]  王沛珍. Thanatin的原核表达、纯化及其抗菌活性的检测[D]. 广州:南方医科大学, 2010.
[24]  Ma YS, Cao W, Zhou YX, et al. Expression and purifcation of antimicrobial peptide adenoregulin with C-amidated terminus in Escherichia coli[J]. Protein Expr Purif, 2005, 40(2):404-410.
[25]  王增, 马会勒, 张文, 等. 包涵体蛋白的分离和色谱法体外复性纯化研究进展[J]. 中国生物工程杂志, 2009, 29(7):102-107.
[26]  冯云, 杨云霞, 黄宁, 等. 人抗菌肽FALL-39在大肠杆菌中高效表达[J]. 生物医学工程杂志, 2003, 20(4):634-637.
[27]  Li Y, Li X, Wang G. Cloning, expression, isotope labeling, and purification of human antimicrobial peptide LL-37 in Escherichia coli for NMR studies[J]. Protein Expr Purif, 2006, 47(2):498-505.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133