劉世強(qiáng); 郭麗瓊; 林俊芳; 郭安平
中國(guó)熱帶農(nóng)業(yè)科學(xué)院熱帶作物生物技術(shù)國(guó)家重點(diǎn)實(shí)驗(yàn)室; 中國(guó)熱帶農(nóng)業(yè)科學(xué)院熱帶作物生物技術(shù)國(guó)家重點(diǎn)實(shí)驗(yàn)室; 中國(guó)熱帶農(nóng)業(yè)科學(xué)院熱帶作物生物技術(shù)國(guó)家重點(diǎn)實(shí)驗(yàn)室; 中國(guó)熱帶農(nóng)業(yè)科學(xué)院熱帶作物生物技術(shù)國(guó)家重點(diǎn)實(shí)驗(yàn)室 海口571701 ; 海口571701; 華南農(nóng)業(yè)大學(xué)食品學(xué)院生物工程系; 廣州510640 ; 海口571701; 華南農(nóng)業(yè)大學(xué)食品學(xué)院生物工程系; 廣州510640 ; 海口571701
【中文摘要】 從金針菇(Flammulina velutipes)7個(gè)菌株中篩選出1株高產(chǎn)植酸酶菌株。根據(jù)GenBank中植酸酶基因的保守區(qū)設(shè)計(jì)并合成一對(duì)特異性引物,以金針菇菌絲的總DNA為模板,通過PCR擴(kuò)增,獲得了一條長(zhǎng)約790bp的片段。DNA序列測(cè)定結(jié)果表明,該片段長(zhǎng)度為788bp,采用Blast軟件進(jìn)行序列比對(duì)發(fā)現(xiàn),該片段與平田頭菇(Agrocybe pediades)的植酸酶基因phy(GenBankAccession:CAC48160)編碼的氨基酸具有64%的序列同源性。該片段含有2個(gè)內(nèi)含子,含有植酸酶基因的活性位點(diǎn)高度保守序列(Active-site sequence)-RHGXRXPT[1]。
【英文摘要】 One Flammulina velutipes strain producing high yield of phytase was screened from seven Flammulina velutipes strains.The phytase gene region was cloned by PCR amplification from Flammulina velutipes genomic DNA with a pair of specific primers designed according to the conserved sequence of phytase genes in the GenBank.DNA sequencing determination results showed that the fragment length was 788bp.Sequence analysis with Blast software indicated that the deduced amino acid sequence was 64% of homology with the amino acid sequence of phytase gene phy(GenBank Accession:CAC48160) from Agrocybe pediades.The fragment contained two introns and the active-site high-degree conserved sequence of phytase genes-RHGXRXPT.
【中文關(guān)鍵詞】 金針菇; 植酸酶; 篩選; 克隆
【英文關(guān)鍵詞】 Flammulina velutipes; Phytase; Screen; Clone
【基金】國(guó)家自然科學(xué)基金資助項(xiàng)目“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)的部分研究?jī)?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.0.2005-03-000