山东农业科学
山東農業科學
산동농업과학
Shandong Agricultural Sciences
2015年
9期
1-6
,共6页
禾谷镰刀菌%14α脱甲基酶(CYP51)%基因克隆%致病性%生物信息学分析
禾穀鐮刀菌%14α脫甲基酶(CYP51)%基因剋隆%緻病性%生物信息學分析
화곡렴도균%14α탈갑기매(CYP51)%기인극륭%치병성%생물신식학분석
Fusarium graminearum%Sterol 14α-demethylation(CYP51)%Gene clone%Pathogenicity%Bioinformatics analysis
本研究采用 RT －PCR 技术，克隆获得禾谷镰刀菌（Fusarium graminearum）致病性 CYP51C 蛋白基因 cDNA 序列，并对蛋白氨基酸序列进行生物信息学分析，明确其序列典型特征。结果表明：其 cDNA 序列全长为1554 bp，编码517个氨基酸；该蛋白分子量为58．6 kD，等电点为6．36；含有细胞色素 P450家族成员典型的保守结构域；不具有信号肽，属于非分泌性蛋白；具有跨膜结构域，是亲水性蛋白；蛋白质二级结构最主要的结构元件是α螺旋和无规则卷曲；亚细胞定位预测显示 CYP51C 蛋白主要位于细胞质中。这些研究结果为蛋白纯化及其蛋白结构生物学研究提供参考，进而可为病原真菌药物靶标设计奠定基础。
本研究採用 RT －PCR 技術，剋隆穫得禾穀鐮刀菌（Fusarium graminearum）緻病性 CYP51C 蛋白基因 cDNA 序列，併對蛋白氨基痠序列進行生物信息學分析，明確其序列典型特徵。結果錶明：其 cDNA 序列全長為1554 bp，編碼517箇氨基痠；該蛋白分子量為58．6 kD，等電點為6．36；含有細胞色素 P450傢族成員典型的保守結構域；不具有信號肽，屬于非分泌性蛋白；具有跨膜結構域，是親水性蛋白；蛋白質二級結構最主要的結構元件是α螺鏇和無規則捲麯；亞細胞定位預測顯示 CYP51C 蛋白主要位于細胞質中。這些研究結果為蛋白純化及其蛋白結構生物學研究提供參攷，進而可為病原真菌藥物靶標設計奠定基礎。
본연구채용 RT －PCR 기술，극륭획득화곡렴도균（Fusarium graminearum）치병성 CYP51C 단백기인 cDNA 서렬，병대단백안기산서렬진행생물신식학분석，명학기서렬전형특정。결과표명：기 cDNA 서렬전장위1554 bp，편마517개안기산；해단백분자량위58．6 kD，등전점위6．36；함유세포색소 P450가족성원전형적보수결구역；불구유신호태，속우비분비성단백；구유과막결구역，시친수성단백；단백질이급결구최주요적결구원건시α라선화무규칙권곡；아세포정위예측현시 CYP51C 단백주요위우세포질중。저사연구결과위단백순화급기단백결구생물학연구제공삼고，진이가위병원진균약물파표설계전정기출。
In this study,the full -length cDNA sequence of pathogenic protein CYP51C from Fusarium graminearum was cloned using RT -PCR,and the bioinformatics analysis of amino acid sequences was con-ducted and its typical characteristics was identified.The results showed that the full -length cDNA sequence of CYP51C included a complete open reading frame (ORF)of 1 554 bp,which encoded 517 amino acids.Its molecular weight was 58.6 kD and its isoelectric point was 6.36.It possessed the conserved domains which were characteristics of cytochrome P450 family members.It was not a secretory protein without a signal pep-tide,but was a hydrophilicity protein with transmembrane domain.The major structural elements of polypep-tide chain were α-helix and random coil.Subcellular localization result indicated that CYP51C was mainly located in cytoplasm.These results would provide references for the study on protein purification and protein structural biology,and then lay a foundation for drug design targeting pathogenic fungi.
