發(fā)布時(shí)間：2018-07-31 20:21

【摘要】：植物脂酰-�；d體蛋白硫酯酶A(fatty acyl acyl carrier thioesterase,FATA)在脂肪酸合成代謝過程中具有重要的調(diào)控作用。目前有關(guān)黃腫樹(Jatropha curcas)脂酰-ACP硫酯酶A基因(JcFATA)和其啟動(dòng)子的表達(dá)模式均還未見報(bào)道。為了研究JcFATA基因的表達(dá)模式,本研究利用半定量RT-PCR和實(shí)時(shí)熒光定量(qRT-PCR)檢測(cè)了JcFATA基因的時(shí)空表達(dá)特異性。結(jié)果表明,JcFATA基因在黃腫樹的多個(gè)組織部位都有表達(dá),尤其是在根、花和葉中表達(dá)量較高。為了分析JcFATA基因啟動(dòng)子的組織表達(dá)模式,本研究將克隆的JcFATA基因啟動(dòng)子片段(PJcFATA)插入到雙元表達(dá)載體pCAMBIA1300G中,成功構(gòu)建了JcFATA基因啟動(dòng)子驅(qū)動(dòng)的β-葡糖醛酸酶(β-glucuronidase,GUS)基因融合表達(dá)載體(p1300G-PJcFATA-GUS);通過農(nóng)桿菌(Agrobacterium tumefaciens)介導(dǎo)的轉(zhuǎn)化方法將重組質(zhì)粒轉(zhuǎn)入野生型擬南芥(Arabidopsis thaliana),經(jīng)過潮霉素篩選,最后對(duì)抗性苗進(jìn)行PCR檢測(cè)和GUS組織化學(xué)分析。PCR檢測(cè)結(jié)果表明,JcFATA基因啟動(dòng)子片段已經(jīng)成功整合到擬南芥基因組中(JGD登錄號(hào):Jcr4S00539)。對(duì)不同發(fā)育時(shí)期的擬南芥植株進(jìn)行GUS組織化學(xué)染色分析,結(jié)果顯示GUS活性具有明顯的時(shí)空特異性,在根、花和葉中的染色較深,有很強(qiáng)的GUS活性。本研究結(jié)果為進(jìn)一步研究JcFATA基因及其啟動(dòng)子功能提供了理論依據(jù)。
[Abstract]:A (fatty acyl carrier thioesterase (A (fatty acyl carrier thioesterase) plays an important role in fatty acid synthesis and metabolism. Up to now, the expression patterns of (JcFATA) and its promoter of (Jatropha curcas) lipoacyl-ACP thioesterase A gene have not been reported. In order to study the expression pattern of JcFATA gene, the temporal and spatial expression specificity of JcFATA gene was detected by semi-quantitative RT-PCR and real-time fluorescence quantification (qRT-PCR). The results showed that JcFATA gene was expressed in many tissues, especially in roots, flowers and leaves. In order to analyze the tissue expression pattern of JcFATA gene promoter, the cloned JcFATA gene promoter fragment (PJcFATA) was inserted into the binary expression vector pCAMBIA1300G. The fusion expression vector (p1300G-PJcFATA-GUS) of 尾 -glucuronidase Gus (尾 -glucuronidase Gus) driven by JcFATA gene promoter was successfully constructed, and the recombinant plasmid was transformed into wild type Arabidopsis thaliana (Arabidopsis thaliana), through hygromycin screening by means of Agrobacterium tumefaciens (Agrobacterium tumefaciens) mediated transformation. The results of PCR and GUS histochemical analysis showed that the promoter fragment of JcFATA gene had been successfully integrated into Arabidopsis thaliana genome (JGD accession number: Jcr4S00539). The results of GUS histochemical staining of Arabidopsis thaliana plants at different developmental stages showed that the activity of GUS had obvious space-time specificity and had strong GUS activity in roots, flowers and leaves. The results provide a theoretical basis for further study of JcFATA gene and its promoter function.
【作者單位】： 廣東海洋大學(xué)農(nóng)學(xué)院;嘉應(yīng)學(xué)院醫(yī)學(xué)院;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(No.31271759) 湛江市科技攻關(guān)項(xiàng)目(No.2016B101)
【分類號(hào)】：Q943.2;S794.9

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