东北林业大学学报
東北林業大學學報
동북임업대학학보
JOURNAL OF NORTHEAST FORESTRY UNIVERSITY
2013年
7期
129-133
,共5页
偏肿革裥菌%锰过氧化物酶%构巢曲霉%原生质体
偏腫革裥菌%錳過氧化物酶%構巢麯黴%原生質體
편종혁간균%맹과양화물매%구소곡매%원생질체
Lenzites gibbosa%Manganese peroxidase (MnP)%Aspergillus nidulans%Protoplast
对已经构建好的携带有白腐菌偏肿革裥菌（Lenzites gibbosa）锰过氧化物酶完整编码序列基因的载体质粒pMDTM18-T/Lg-mnp、以及整合型表达载体质粒pLB01分别双酶切，然后进行连接构建了重组质粒pLB01/Lg-mnp。对构巢曲霉（Aspergillus nidulans）尿嘧啶尿苷营养缺陷体菌株TN02A7进行了制备分生孢子原生质体酶解方法的摸索。结果表明：将溶壁酶、纤维素酶和蜗牛酶3种酶以1∶1∶1的比例混合，能有效地使构巢曲霉的分生孢子形成原生质体。采用PEG/CaCl2介导的原生质体转化方法成功地将L．gibbosa的MnP基因转入到了构巢曲霉中，获得了携带有白腐菌基因的构巢曲霉转化子菌株。
對已經構建好的攜帶有白腐菌偏腫革裥菌（Lenzites gibbosa）錳過氧化物酶完整編碼序列基因的載體質粒pMDTM18-T/Lg-mnp、以及整閤型錶達載體質粒pLB01分彆雙酶切，然後進行連接構建瞭重組質粒pLB01/Lg-mnp。對構巢麯黴（Aspergillus nidulans）尿嘧啶尿苷營養缺陷體菌株TN02A7進行瞭製備分生孢子原生質體酶解方法的摸索。結果錶明：將溶壁酶、纖維素酶和蝸牛酶3種酶以1∶1∶1的比例混閤，能有效地使構巢麯黴的分生孢子形成原生質體。採用PEG/CaCl2介導的原生質體轉化方法成功地將L．gibbosa的MnP基因轉入到瞭構巢麯黴中，穫得瞭攜帶有白腐菌基因的構巢麯黴轉化子菌株。
대이경구건호적휴대유백부균편종혁간균（Lenzites gibbosa）맹과양화물매완정편마서렬기인적재체질립pMDTM18-T/Lg-mnp、이급정합형표체재체질립pLB01분별쌍매절，연후진행련접구건료중조질립pLB01/Lg-mnp。대구소곡매（Aspergillus nidulans）뇨밀정뇨감영양결함체균주TN02A7진행료제비분생포자원생질체매해방법적모색。결과표명：장용벽매、섬유소매화와우매3충매이1∶1∶1적비례혼합，능유효지사구소곡매적분생포자형성원생질체。채용PEG/CaCl2개도적원생질체전화방법성공지장L．gibbosa적MnP기인전입도료구소곡매중，획득료휴대유백부균기인적구소곡매전화자균주。
We digested the integrated expression vectors pLB01 and pMDTM 18-T/Lg-mnp by restriction endonuclease XbaI and BamHI, respectively, and then connected them by T4 DNA ligase to construct the recombinant plasmid pLB01/Lg-mnp. We explored the methods how the conidiophores of auxotrophic stain TN02A7 of Aspergillus nidulans were transformed to protoplasts by different cell wall lyases.Lywallzyme, cellulase and snailase mixed together by 1 ∶1 ∶1 can effectively make the spores become protoplasts.By PEG/CaCl2 mediated protoplast transformation method, a manganese peroxidase gene of Lenzites gibbosa is successfully transferred to TN02A7.
