枯草芽孢桿菌精氨酸脫羧酶基因speA的表達(dá)與蛋白純化
發(fā)布時(shí)間:2018-03-12 09:01
本文選題:枯草芽孢桿菌 切入點(diǎn):精氨酸脫羧酶 出處:《中國釀造》2017年03期 論文類型:期刊論文
【摘要】:根據(jù)枯草芽孢桿菌(Bacillus subtilis)BJ3-2的精氨酸脫羧酶(ADC)的編碼基因spe A序列設(shè)計(jì)特異性酶切引物,克隆基因speA序列。測序結(jié)果顯示,基因speA全長為1473 bp,編碼490個(gè)氨基酸,分子質(zhì)量為58 ku。基因spe A克隆至原核表達(dá)載體,獲得重組菌pET28a-spe A/BL21,十二烷基硫酸鈉聚丙烯酰胺凝膠電泳(SDS-PAGE)結(jié)果顯示,1.0 mmol/L的異丙基β-D-硫代半乳糖苷(IPTG)28℃誘導(dǎo)4 h,上清液和菌體均能表達(dá)出ADC蛋白,上清液經(jīng)純化、透析、冷凍干燥可獲得純度97%的ADC酶,酶聯(lián)免疫吸附檢測(ELISA)ADC酶活為16780 U/mg。為speA基因的表達(dá)、純化及酶學(xué)性質(zhì)研究奠定了理論基礎(chǔ)。
[Abstract]:According to the speA sequence of the arginine decarboxylase of Bacillus subtilis)BJ3-2, the specific primers were designed and the speA sequence of the gene was cloned. The results showed that the length of the gene speA was 1473 BP, encoding 490 amino acids. The molecular weight was 58 ku. the gene spe A was cloned into prokaryotic expression vector. The results of SDS-PAGE showed that the isopropyl 尾 -D- thiogalactoside (IPTG) of 1.0 mmol/L was induced at 28 鈩,
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