阔褶水蛙皮肤组织cDNA文库的构建及抗菌肽brevinin-2LTs的cDNA克隆和序列分析

doi:10.7668/hbnxb.2009.S1.003

华北农学报

阔褶水蛙皮肤组织cDNA文库的构建及抗菌肽brevinin-2LTs的cDNA克隆和序列分析

王辉¹, 张秀清¹, 胡玉红², 刘敬兰², 孙君社¹

1. 中国农业大学食品科学与营养工程学院,北京100083;
2. 河北师范大学实验中心,河北石家庄050016

收稿日期:2009-01-15 出版日期:2009-12-31
通讯作者: 孙君社(1961-),男,河北栾城人,博士生导师,主要从事生物技术在农产品加工中应用和酶与发酵方面的教学与研究工作。
作者简介:王辉(1981-),男,博士,主要从事蛙类功能性多肽研究。
基金资助:
河北师范大学青年基金(L2007Q32)

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

WANGHui¹, ZHANGXiu-qing¹, HUYu-hong², LIUJing-lan², SUNJun-she¹

1. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China;
2. Experimental Center, Hebei Normal University, Shijiazhuang 050016, China

Received:2009-01-15 Published:2009-12-31

摘要/Abstract

摘要： 运用SMART技术成功构建了阔褶水蛙(Hylaranalatouchii)皮肤组织全长cDNA文库,用来筛选抗菌肽及其他功能性基因。经鉴定,原始文库滴度为9.7×106cfu/mL,重组率达到91%,插入片段大小在0.3~1.5kb范围内,文库扩增后的滴度为1.9×109cfu/mL,说明所构建文库质量较高。通过该文库成功筛选出2个抗菌肽cDNA序列,此序列属于brevinin-2家族,因此命名为brevinin-2LT1和brevinin-2LT2。

关键词: 阔褶水蛙, cDNA文库, 抗菌肽, 文库滴度

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

中图分类号:

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.

http://www.hbnxb.net/CN/Y2009/V24/IS1/9

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