季哲; 李玉祥; 趙明文; 潘迎捷
南京農(nóng)業(yè)大學(xué)農(nóng)業(yè)部農(nóng)業(yè)環(huán)境微生物工程重點(diǎn)開(kāi)放實(shí)驗(yàn)室; 南京農(nóng)業(yè)大學(xué)農(nóng)業(yè)部農(nóng)業(yè)環(huán)境微生物工程重點(diǎn)開(kāi)放實(shí)驗(yàn)室; 南京農(nóng)業(yè)大學(xué)農(nóng)業(yè)部農(nóng)業(yè)環(huán)境微生物工程重點(diǎn)開(kāi)放實(shí)驗(yàn)室; 上海水產(chǎn)大學(xué) 南京210095 ; 南京210095 ; 南京210095 ; 上海200090
【中文摘要】 本研究以黃傘菌絲cDNA為模板,設(shè)計(jì)簡(jiǎn)并引物,經(jīng)PCR擴(kuò)增出兩條片段SL1和SL2,經(jīng)過(guò)回收、重?cái)U(kuò)增、克隆與鑒定后進(jìn)行測(cè)序分析,結(jié)果發(fā)現(xiàn)SL1的序列在Genebank中與三磷酸甘油醛脫氫酶(Glyceraldehyde-3-phosphate dehydrogease,GPD)基因的同源性達(dá)到85%,根據(jù)SL1序列設(shè)計(jì)引物,以黃傘菌絲體、原基、菇蕾、子實(shí)體菌柄、子實(shí)體菌蓋的cDNA為模板進(jìn)行PCR反應(yīng)以驗(yàn)證其保守性,結(jié)果顯示此片段在不同分化發(fā)育階段表達(dá)量相同。
【英文摘要】 The mixed primer was designed,and two fragments SL1 and SL2 were acquired by PCR templating of the cDNA of Pholiota adiposa(Fr.)quél mycelium.After reamplifying,purifying,cloning,measuring sequence and blasting in GeneBank,the result showed that identified SL1 fragment is similar to Glyceraldehyde-3-phosphate dehydrogease(GPD)gene,and the homology is as high as 85%.The primer is designed on the basis of SL1,cDNA of different period including mycelium,anlage,bud and fruit body(stem and lid of mushroom)was extracted and treated by PCR to verifying the conservativeness.The result shows that the expression quantity of this fragment is uniform during different developing period of Ph adipose.
【中文關(guān)鍵詞】 黃傘; 三磷酸甘油醛脫氫酶基因; 克隆
【英文關(guān)鍵詞】 Pholiota adiposa; Glyceraldehyde-3-phosphate dehydrogease(GPD)gene; Cloning
【文獻(xiàn)出處】 中國(guó)食用菌,Edible Fungi of China,編輯部郵箱,2005年05期 【DOI】CNKI:SUN:ZSYJ.0.2005-05-019