摘要
为克隆和研究软腐病菌Dickeya dadantii的信号分子合成酶基因expI,从Dickeya属模式菌DSM–18020的基因组中扩增获得了基因expI。生物信息学分析显示,该基因全长639 bp,与Dickeya dadantii 3937的N–酰基高丝氨酸内酯(N–acyl–homoserine lactone,AHLs)合成酶基因同源性达99%;DSM–18020 expI编码的蛋白质相对分子质量为24 704,理论等电点为5.85,整个肽链中均匀分布疏水氨基酸,且比亲水性氨基酸多,没有信号肽存在;将获得的exp I基因构建到表达载体p ET–28α(+)并转入大肠杆菌BL21(DE3),诱导后的表达产物与理论值一致。
In order to study the signal molecule synthase gene expI of Dickeya dadantii, the gene expI was amplified from the genome of the type strain DSM–18020. Bioinformatics analysis showed that there was a complete open reading frame(ORF) in DSM–18020 expI, which was 639 bp in length and had 99% homology to N–acyl–homoserine lactone(AHLs) synthetase of D. dadantii 3937. The molecular weight of the protein encoded by DSM–18020 expI is 24,704, and the theoretical isoelectric point is 5.85. The hydrophobic amino acids are more abundant than hydrophilic amino acids and distributed evenly throughout the peptide chain, with no signal peptide. expI gene was cloned into the expression vector p ET28α and transformed into Escherichia coli BL21(DE3). The expression product was basically identical with the expectation.
引文
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