CAJ | 학술논문

克隆渐狭蜡蚧菌几丁质酶基因，为进一步明确该菌产生的几丁质酶对定居性根结类和孢囊类线虫卵孵化抑制作用提供理论依据。采用分离自黑龙江大豆孢囊线虫孢囊寄生真菌CGMCC5328，通过形态学特征及ITS序列比较分析，鉴定该菌株为渐狭蜡蚧菌。通过简并引物设计和RACE技术，克隆渐狭蜡蚧菌中几丁质酶基因，利用生物学软件分析该基因序列及其编码的氨基酸序列。首次从渐狭蜡蚧菌中克隆得到一个几丁质酶基因LACHI1，该基因DNA序列长1743 bp，含3个内含子，包含1272 bp开放阅读框，编码423个氨基酸，理论分子量45．9 kDa，等电点5.90。 LACHI1基因编码的几丁质酶，属于糖基水解酶18家族几丁质酶。同源性比对表明该菌产生的几丁质酶与昆虫寄生真菌和食线虫真菌产生的几丁质酶有很高的同源性。
극륭점협사개균궤정질매기인，위진일보명학해균산생적궤정질매대정거성근결류화포낭류선충란부화억제작용제공이론의거。채용분리자흑룡강대두포낭선충포낭기생진균CGMCC5328，통과형태학특정급ITS서렬비교분석，감정해균주위점협사개균。통과간병인물설계화RACE기술，극륭점협사개균중궤정질매기인，이용생물학연건분석해기인서렬급기편마적안기산서렬。수차종점협사개균중극륭득도일개궤정질매기인LACHI1，해기인DNA서렬장1743 bp，함3개내함자，포함1272 bp개방열독광，편마423개안기산，이론분자량45．9 kDa，등전점5.90。 LACHI1기인편마적궤정질매，속우당기수해매18가족궤정질매。동원성비대표명해균산생적궤정질매여곤충기생진균화식선충진균산생적궤정질매유흔고적동원성。
To clone chitinolytic genes from Lecanicillium attenuatum and provide theoretical basis for the inhibi-tion of the chitinases on egg-hatching of root knot and cyst nematode .In this study ,an nematode egg-parasitic fungus CGMCC5328 was isolated from Heterodera glycines infecting soybean in Heilongjiang Province .Based on morphological characters and molecular analysis of ITS-rDNA,the strain was identified as L.attenuatum.The chitinase gene LACHI1 from L.attenuatum was cloned using the degenerate PCR primers and RACE techniques .The analysis of the gene LA-CHI1 and its amino acid sequence was by biology software .We have cloned a chitinase gene LACHI1 from L.attenua-tum for the first time.The gene is 1 743 bp in length and contains three putative introns .The ORF of LACHI1 is 1 272 bp in size with encoding protein of 423 aa,molecular mass of 45.9 kDa and pI of 5.90.The chitinase deduced by LA-CHI1 belongs to glycosyl hydrolase family 18 chitinase .Comparison of the chitinase amino acid sequence with other chitinases from entomopathogenic fungi and nematophagous fungi revealed that the enzymes were highly similar .