棉花UXS基因生物信息学分析与原核表达

Bioinformatic analysis and prokaryotic expression of three UXS genes in cotton(Gossypium hirsutum)

通过生物信息学方法对已克隆的3个棉花UXS基因GhUXS1、GhUXS2和GhUXS3编码产物进行亲疏水性,结构域以及功能等多方面预测,同时将产物连接至pET-32a(+)原核表达载体,并转化BL21(DE3)plys菌株,在IPTG诱导下,利用Western blotting方法检测GhUXS1、GhUXS2和GhUXS3在原核细胞中的表达。生物信息学预测发现3个蛋白均含有UXS家族的保守域。GhUXS1和GhUXS2结构类似,N端含有较为明显的疏水区以及跨膜区,GhUXS3N端缺乏明显疏水区及跨膜区。Western blotting结果显示在体外分别成功表达GhUXS1、GhUXS2和GhUXS3和His标签的融合蛋白。

Coding products of three cotton UXS（UDP-glucuronate decarboxylase） genes,GhUXS1,GhUXS2 and GhUXS3,were predicted with bioinformatics methods,including hydrophilic-hydrophobic property,domains and functions.Then three coding sequences were cloned into pET-32a（＋） prokaryotic expression vector and transformed into BL21（DE3） plus strain.After corresponding fusion proteins being induced by IPTG,Western blotting was then employed to detect their expressions in prokaryotic cells.The result showed that all three proteins had conserved domain of NAD-binding in UXS family.GhUXS1 and GhUXS2 had similar structure,for example,with obvious hydrophobic region and transmembrane region in N-terminal,while GhUXS3 lacked those structures.Western blotting results indicated that all three fusion proteins were successfully expressed.