摘要
侧孢短芽孢杆菌(Brevibacillus laterosporus)S62-9菌株产生的抗菌肽BBL属于非核糖体合成肽,其生物合成涉及到一系列相关基因。为了获得完整的抗菌肽BBL基因决定簇,本研究基于部分已知的NRPS-A结构域基因序列,以S62-9菌株质粒DNA为模板,采用TAIL-PCR技术对NRPS-A结构域的上游进行扩增,得到一个大小为2.74kb的序列,该序列包含NRPS-A的5′末端序列、pp-binding序列以及1个ABC转运蛋白基因。该序列进一步与已知序列进行序列拼接及软件分析比对,最终获得一个大小为22.2kb的基因簇,该基因簇包含13个Bbl功能结构域,根据功能该基因簇分为3类功能相关基因:一类是编码转运相关蛋白,BblE包含3个ABC转运蛋白,负责抗菌肽向胞外渗透运输;二是催化短肽合成基因,Bbl F包含了pp-binding结构域和NRPS-A结构域,与氨基酸的装载和缩合有关,负责短肽的合成;三是肽链修饰相关基因,Bbl D、Bbl E、Bbl M、Bbl K等基因负责肽的修饰。
Antimicrobial peptide BBL produced by Brevibacillus laterosporus S62 9 strain belongs to non-ribosomal synthesis peptide,the synthesis of which involves a range of related genes.In order to get the complete antimicrobial peptide gene cluster,the primers were designed based on the partially known NRPS-A domain gene sequence,and TAIL-PCR was carried out using plasmid DNA of S62 9 as a template.The results showed that a length of 2.74 kb sequence was amplified,containing 5′-ter minal sequence of NRPS-A,the pp-binding sequence and one ABC transporter.This sequence was further aligned with the known sequence and analyzed by software,and a size of 22.2 kb gene cluster containing 13 Bbl domains was obtained,which was classified into 3 functional groups:groupⅠ(Bbl E)encoded the related transport proteins,containing 3 ABC transporters,which were responsible for transmembrane transport of BBL;groupⅡ(Bbl F)included a PP-binding domain and a NRPS-A domain,and were responsible for the synthesis of BBL;groupⅢ(Bbl D、Bbl E、Bbl M、Bbl K Etc.)played an important role in the modification for BBL.
作者
刘洋
柯晓静
于宏伟
郭润芳
LIU Yang;KE Xiaojing;YU Hongwei;GUO Runfang(College of Food Science and Technology,Hebei Agricultural University,Baoding 071000,China)
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2019年第1期65-70,共6页
Journal of Hebei Agricultural University
基金
国家科技支撑计划(2013BAD10B03)
河北省自然科学基金(C2019204043)
关键词
非核糖体肽合成酶
交错式热不对称
克隆
结构特征
non-ribosomal peptide synthetase
thermal asymmetric interlaced PCR
cloning
structural characteristics
