本文選題：參環(huán)毛蚓　切入點(diǎn)：重金屬富集　出處：《中草藥》2016年16期 　論文類型：期刊論文

【摘要】：目的揭示參環(huán)毛蚓MT-2基因的轉(zhuǎn)錄調(diào)控機(jī)制,為今后采用基因工程技術(shù)改良參環(huán)毛蚓重金屬富集性狀奠定基礎(chǔ)。方法根據(jù)已知的MT-2c DNA序列,設(shè)計(jì)特異引物,通過(guò)PCR擴(kuò)增參環(huán)毛蚓MT-2基因的編碼區(qū)基因組序列,擴(kuò)增產(chǎn)物經(jīng)測(cè)序后自行比對(duì)分析�；谠撔蛄性O(shè)計(jì)3條特異引物,采用基因步移技術(shù)克隆其上游的啟動(dòng)子序列,同時(shí)運(yùn)用Promoter Prediction在線軟件分析預(yù)測(cè)其順式元件。結(jié)果經(jīng)PCR擴(kuò)增、測(cè)序后獲得2 826 bp MT-2基因編碼區(qū)序列,將該序列與已知MT-2 c DNA序列(登錄號(hào)為KC787373.1)進(jìn)行比對(duì),發(fā)現(xiàn)MT-2編碼區(qū)由4個(gè)外顯子和4個(gè)內(nèi)含子組成�；虿揭萍夹g(shù)克隆得到1 534 bp的啟動(dòng)子序列,經(jīng)Promoter Prediction在線軟件分析,結(jié)果顯示該序列不僅含有TATA-box、CAAT-box等核心啟動(dòng)子軟件,還具有3個(gè)特異響應(yīng)重金屬參與調(diào)控MT-2表達(dá)的MRE元件。結(jié)論參環(huán)毛蚓的MT-2基因能被重金屬誘導(dǎo)表達(dá),MT-2基因轉(zhuǎn)錄水平的金屬調(diào)控是通過(guò)MT-2基因啟動(dòng)子區(qū)的金屬調(diào)控元件MRE來(lái)實(shí)現(xiàn)的。
[Abstract]:Objective to elucidate the transcriptional regulation mechanism of MT-2 gene and lay a foundation for the improvement of heavy metal enrichment by genetic engineering. Methods based on the known MT-2c DNA sequence, specific primers were designed. The coding region of MT-2 gene was amplified by PCR, and the amplified product was sequenced and analyzed. Based on the sequence, three specific primers were designed, and the upstream promoter sequence was cloned by gene step technique. At the same time, the cis element was predicted by Promoter Prediction online software. Results after PCR amplification, the sequence of 2826 BP MT-2 gene coding region was obtained, and the sequence was compared with the known MT-2 c DNA sequence (accession number KC787373.1). It was found that the coding region of MT-2 was composed of 4 exons and 4 introns. The promoter sequence of 1 534 BP was cloned by gene step technique and analyzed by Promoter Prediction online software. The results showed that the sequence contained not only the core promoter software such as TATA-box CAAT-box, but also the core promoter software such as TATA-box. There are also three MRE elements that specifically respond to heavy metals to regulate the expression of MT-2. Conclusion the metal regulation of the transcription level of the MT-2 gene can be induced by heavy metal in the promoter region of the MT-2 gene through the metal regulation of the promoter region of the MT-2 gene. Component MRE to achieve.
【作者單位】： 廣州中醫(yī)藥大學(xué)中藥學(xué)院;湖北中醫(yī)藥大學(xué)藥學(xué)院;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(30772741) 廣東省科技計(jì)劃項(xiàng)目(2009B060700006) 廣州市科技計(jì)劃項(xiàng)目(2010Y1-C981)
【分類號(hào)】：S865.4;Q78

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