Construction of cDNA Library of Hylaranala touchi Skin Tissue with Cloningand Sequence Analysis of the cDNA Encoding Antimicrobial Peptides Brevinin-2LTs

doi:10.7668/hbnxb.2009.S1.003

Abstract

Abstract: A full length cDNA library of Hylarana latouchii skin tissue was constructed with the SMART technique.This cDNA library could be used to screen encoding gene of antimicrobial peptides and other funct ional genes.The titer ofthe cDNA library was 9. 7 @ 106 cfu/ mL, and the recombination rate of it was 91% . Inserted fragments ranged was 013-115 kb.The titer of the amplified library was 119 @ 109 cfu/ mL. Successfully constructed cDNA library was a full lengthlibrary with high quality.Two different cDNA sequences encoding antimicrobial peptide precursors named brevinin- 2LT1and brevinin - 2LT2 were cloned by PCR from this cDNA library. All of the two antimicrobial pept ides attributed tobrevinin- 2 family.

Key words: HylaranaLatouchii, cDNAlibrary, Antimicrobialpeptides, Titer

CLC Number:

Q785

WANGHui, ZHANGXiu-qing, HUYu-hong, LIUJing-lan, SUNJun-she. Construction of cDNA Library of Hylaranala touchi Skin Tissue with Cloningand Sequence Analysis of the cDNA Encoding Antimicrobial Peptides Brevinin-2LTs[J]. ACTA AGRICULTURAE BOREALI-SINICA, doi: 10.7668/hbnxb.2009.S1.003.

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References

[1] 赖仞,冉永禄. 两栖类皮肤活性肽与活性生物胺[J].大自然探索,1999,18( 67):71- 74.
[2] Ganz T. Defensins: antimicrobial peptides of innate immunity[J].Nat Rev Immunol,2003,3( 9):710- 720.
[3] Skerlavaj B,Benincasa M,Risso A,et al.SMAP- 29: A potentantibacterial and antifungal peptide from sheep leucocytes[J].FEBS Lett,1999,463( 1- 2):58- 62.
[4] Wang H X,Ng T B. Isolation and characterization of an ant-ifungal peptide with antiproliferative activity from seeds ofPhaseolus vulgaris cv.c Spotted Beanc [J ].Appl MicrobiolBiotechnol,2007,74( 1):125- 130.
[5] Wu S,Jia S,Sun D,et al. Pur ification and characterization oftwo novel antimicrobial peptides Subpeptin JM4- A and subpeptinJM4- B produced by Bacillus subtilis JM4 [J].CurrMicrobiol,2005,51( 5):292- 296.
[6] Zhou J,McClean S,Thompson A,et al.Purification and characterizationof novel antimicrobial peptides from the skin secretionof Hylarana guentheri [J].Peptides,2006,27( 12) :3077- 3084.
[7] Chen T,Li L,Zhou M,et al. Amphibian skin peptides andtheir corresponding cDNAs from single lyophilized secretionsamples: identification of novel brevinins from three species ofChinese frogs[J].Peptides,2006,27( 1):42- 48.
[8] Suzuki H,Iwamuro S,Ohnuma A,et al.Expression of genesencoding antimicrobial and bradykinin - related peptides inskin of the stream brown frog Rana sakuraii [J].Peptides,2007,28( 3):505- 514.
[9] Conlon JM,Kolodziejek J,Nowotny N. Antimicrobial peptidesfrom ranid frogs: taxonomic and phylogenetic markers and apotential source of new therapeutic agents [J].Biochim BiophysActa,2004,1696( 1):1- 14.
[10] Conlon J M,Sonnevend A,Patel M,et al.A family ofbrevinin - 2 peptides with potent activity against Pseudomonasaeruginosa from the skin of the Hokkaido frog,Rana pirica [J].Regul Pept,2004,118( 3):135- 141.
[11] 邓艳,魏冬霞,吴绍强,等. 猪蛔虫雄虫cDNA 文库的构建[J].畜牧兽医学报,2006,37( 8):804- 808.
[12] 魏冬霞,邓艳,吴绍强,等. 用SMART 技术构建猪蛔虫雌虫cDNA 文库[J].中国预防兽医学报,2007,29( 3):180- 184.

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