γ-聚谷氨酸的基因克隆与序列分析

发布时间：2019-02-17 11:50

【摘要】：本研究通过基因组DNA提取和PCR扩增分别得到Bacillusnatto20646的原初始菌株N-1、高压电晕电场和不同种类稀土元素诱变的突变株N-2、N-3、N-4、N-5及N-6的pgsBCA合成酶基因序列,利用BioEdit软件对6株菌株的pgsB、pgsC和pgsA的基因序列及合成酶蛋白PgsB、PgsC和PgsA蛋白酶的氨基酸序列对比后得出:pgsB、pgsC和pgsA基因分别约含1170个、450个和1140个核苷酸,分别编码约390个、150个和380个氨基酸。高压电晕电场和稀土元素使得菌株γ-PGA合成酶基因簇pgsBCA核苷酸发生了碱基的置换和颠换、插入和缺失,合成酶蛋白PgsBCA氨基酸序列发生了氨基酸的替换、插入和缺失,且对序列的开端和尾端的影响较大,PgsA在30~40个氨基酸区间变化显著,使该处氨基酸残基的跨膜区发生变化,准确的膜锚定,γ-PGA高效地从活性中心位点移开,加快实现了链的延长,为γ-PGA产量的提高提供理论基础。
[Abstract]:In this study, the original strain N-1 of Bacillusnatto20646 was obtained by genomic DNA extraction and PCR amplification, and the mutant N-2ON-3N4 induced by high-voltage electric field and different rare earth elements was obtained, respectively. The sequence of pgsBCA synthase gene of N-5 and N-6 was compared with that of pgsB,pgsC and pgsA of 6 strains by BioEdit software, and the amino acid sequence of PgsB,PgsC and PgsA protease were compared. The results showed that: pgsB, PgsC and pgsA genes contain about 1170 nucleotides 450 nucleotides and 1140 nucleotides encoding 390 150 and 380 amino acids respectively. The high voltage halo electric field and rare earth elements caused the substitution, transversion, insertion and deletion of the nucleotides of the pgsBCA gene cluster of the 纬-PGA synthase gene, and the amino acid substitution, insertion and deletion of the amino acid sequence of the synthase protein PgsBCA. The change of PgsA was significant in the range of 30 ~ 40 amino acids. The transmembrane region of amino acid residues in the region changed, and the accurate membrane anchoring, 纬-PGA effectively removed from the active center site. The chain lengthening is accelerated and the theoretical basis is provided for the increase of 纬-PGA yield.
【作者单位】：内蒙古大学物理科学与技术学院自治区离子束生物工程重点实验室;内蒙古农业大学理学院;
【基金】：国家自然科学基金(51267014)高压电晕电场对微生物诱变效应的研究(30101-131135)资助
【分类号】：Q78

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