于曉玲; 林俊芳; 郭麗瓊; 王樹昌
中國熱帶農(nóng)業(yè)科學(xué)院熱帶作物生物技術(shù)國家重點(diǎn)實(shí)驗(yàn)室; 中國熱帶農(nóng)業(yè)科學(xué)院熱帶作物生物技術(shù)國家重點(diǎn)實(shí)驗(yàn)室; 中國熱帶農(nóng)業(yè)科學(xué)院熱帶作物生物技術(shù)國家重點(diǎn)實(shí)驗(yàn)室; 中國熱帶農(nóng)業(yè)科學(xué)院熱帶作物生物技術(shù)國家重點(diǎn)實(shí)驗(yàn)室 熱帶生物技術(shù)研究所?？?71101 ; 熱帶生物技術(shù)研究所?？?71101; 華南農(nóng)業(yè)大學(xué)食品學(xué)院生物工程系; 廣州510640 ; 熱帶生物技術(shù)研究所?？?71101; 華南農(nóng)業(yè)大學(xué)食品學(xué)院生物工程系; 廣州510640 ; 熱帶生物技術(shù)研究所?？?71101
【中文摘要】 以香菇基因組DNA為模板,利用PCR技術(shù)克隆了香菇1個(gè)ras啟動(dòng)子Lras,2個(gè)gpd啟動(dòng)子Lgpd1和Lgpd2。這3個(gè)啟動(dòng)子的大小分別為715bp、1056bp和611bp,與已報(bào)道的ras和gpd啟動(dòng)子的同源性很強(qiáng)。對這3個(gè)啟動(dòng)子的分析結(jié)果表明,它們都含有豐富的啟動(dòng)子順式作用元件。將這些啟動(dòng)子片段分別與GUS基因連接構(gòu)建了表達(dá)載體,并將這些表達(dá)載體導(dǎo)入草菇菌絲體中,檢測其活性,最終顯示3個(gè)片段都有啟動(dòng)GUS基因表達(dá)的能力,其中Lgpd1啟動(dòng)子活性最強(qiáng),Lras啟動(dòng)子次之,Lgpd2最弱。
【英文摘要】 The promoter fragments of Lras,Lgpd1 and Lgpd2 were isolated from genomic DNA of Lentinus edodes by PCR amplification.DNA sequencing results showed that these promoter fragments have 715bp,1056bp and 611bp respectively.Blastn analysis results revealed that Lras,Lgpd1 and Lgpd2 have high homologous with reported promoters,ras and gpd.Analysis results showed that they all contain many important cis-acting regulatory elements.Fragments of Lras,Lgpd1 and Lgpd2 were constructed respectively with pCAMBIA1301 which 35S promoter was deleted.Then the new expression plasmids containing GUS reporter gene were induced into Volvariella volvacea by protoplast electroporation.GUS histochemistry assay results showed that Lgpd1 had the strongest promoter activity,Lras had stronger activity and Lgpd2 had slight activity.
【中文關(guān)鍵詞】 香菇; ras啟動(dòng)子; gpd啟動(dòng)子; 原生質(zhì)體轉(zhuǎn)化; GUS基因
【英文關(guān)鍵詞】 Lentinus edodes; ras promoter; gpd promoter; Protoplast transformation; GUS
【基金】國家自然科學(xué)基金資助項(xiàng)目“抗冷凍蛋白基因遺傳轉(zhuǎn)化草菇的研究”(編號(hào):39960050);; “草菇冷凍脅迫誘導(dǎo)表達(dá)基因及其啟動(dòng)子的克隆分離”(編號(hào):30060054);; “食用菌外源基因表達(dá)系統(tǒng)的建立”(編號(hào):30371000);; 廣東省自然科學(xué)基金資助項(xiàng)目“食用菌外源基因表達(dá)系統(tǒng)的建立”(編號(hào):032239)的部分研究內(nèi)容
【文獻(xiàn)出處】 食用菌學(xué)報(bào),Acta Edulis Fungi,編輯部郵箱,2005年03期 【DOI】CNKI:SUN:SYJB.0.2005-03-002