本文選題：基因表達 + 谷胱甘肽��； 參考：《基因組學與應(yīng)用生物學》2017年05期

【摘要】：谷胱甘肽(glutathione,GSH)是生物體內(nèi)的非蛋白巰基類化合物,具有抗氧化、抗衰老和重金屬解毒等重要的生理功能。本研究從煙草(Nicotiana tabacum L.)中克隆到谷胱甘肽合成酶基因NtGSH2,該基因編碼555個氨基酸殘基的蛋白質(zhì)。氨基酸序列比對和進化樹分析發(fā)現(xiàn),NtGSH2與其他植物同源蛋白具有較高的一致性,且與枸杞(Lycium chinense)LcGSHS/LcGS和番茄(Solanum lycopersicum)SlGSH2歸為一類,親緣關(guān)系最近。組織表達模式分析表明,NtGSH2基因在煙草的根、莖、葉、花和種子中均表達,其中在花中的表達量最高,在種子中的表達量最低。實時熒光定量PCR檢測表明,NtGSH2受CdCl_2和ZnCl_2誘導(dǎo)表達。將NtGSH2連接到原核表達載體pET32a上,轉(zhuǎn)化獲得含有NtGSH2基因的大腸桿菌BL21(DE3)工程菌并成功表達目的蛋白。該研究結(jié)果為進一步分析NtGSH2基因功能及谷胱甘肽的生物合成提供了一定的理論依據(jù)。
[Abstract]:Glutathione glutathione (GSH) is a nonprotein sulfhydryl compound which has important physiological functions such as antioxidation, anti-aging and detoxification of heavy metals. In this study, Nicotiana tabacum L. The glutathione synthase gene NtGSH _ 2, which encodes 555 amino acid residues, was cloned. Amino acid sequence alignment and phylogenetic tree analysis showed that NtGSH _ 2 had high consistency with other plant homologous proteins, and was closely related to Lycium chinense)LcGSHS/LcGS and Solanum lycopersicum)SlGSH2 of Lycium barbarum. Tissue expression pattern analysis showed that NtGSH2 gene was expressed in roots, stems, leaves, flowers and seeds of tobacco, among which the highest expression was in flowers and the lowest in seeds. Real time fluorescence quantitative PCR assay showed that NtGSH2 was induced by CdCl_2 and ZnCl_2. NtGSH2 was ligated into the prokaryotic expression vector pET32a and transformed into E. coli BL21 (DE3) containing NtGSH2 gene and the target protein was successfully expressed. The results provide a theoretical basis for further analysis of NtGSH2 gene function and glutathione biosynthesis.
【作者單位】： 西南科技大學生命科學與工程學院;西南科技大學核廢物與環(huán)境安全國防重點實驗室;
【基金】：國家核設(shè)施退役及放射性廢物治理科研重點項目(No.14ZG6101) 國家自然科學基金(No.31500205) 西南科技大學核廢物與環(huán)境安全國防重點學科實驗室預(yù)研基金(No.15yyhk09);西南科技大學重點科研平臺專職科研創(chuàng)新團隊基金(No.14tdhk02)共同資助
【分類號】：Q943.2

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1 魏鎖成;張憲云;;谷胱甘肽合成酶的克隆與工程菌篩選[J];科技導(dǎo)報;2008年03期

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