CAJ | 학술논문

目的从马尔尼菲青霉菌(PM)酵母相全长cDNA文库中识别溶血磷脂酶基因,预测其结构和功能,为进一步实验研究提供理论依据.方法利用NCBI在线分析工具对PM酵母相全长cDNA文库进行筛选,识别溶血磷脂酶基因;通过Vector NTI suite 8.0软件包,对其编码蛋白质的各种结构与功能特征进行预测,并构建种系分子进化树.结果筛选得到的基因长1014 bp,Blastx分析该基因可能是编码PM溶血磷脂酶的全长基因,完整的开放读码框(ORF)共含729 bp,编码243个氨基酸;其编码蛋白的相对分子质量为26 800,预测等电点为5.49,疏水氨基酸占47.7%,亲水氨基酸占26.8%,酸性氨基酸占12.7%,碱性氨基酸占12.8%;该蛋白有潜在的2个酪蛋白激酶Ⅱ磷酸化位点、3个蛋白激酶C磷酸化位点和7个N-肉豆蔻酰位点.结论该氨基酸序列属于溶血磷脂酶样功能域特征蛋白酶;与球孢子菌亲源关系最近.通过研究,一个PM溶血磷脂酶基因被成功发现,这为进一步探讨该基因的功能提供了条件.
목적 종마이니비청매균(PM)효모상전장cDNA문고중식별용혈린지매기인,예측기결구화공능,위진일보실험연구제공이론의거.방법 이용NCBI재선분석공구대PM효모상전장cDNA문고진행사선,식별용혈린지매기인;통과Vector NTI suite 8.0연건포,대기편마단백질적각충결구여공능특정진행예측,병구건충계분자진화수.결과 사선득도적기인장1014 bp,Blastx분석해기인가능시편마PM용혈린지매적전장기인,완정적개방독마광(ORF)공함729 bp,편마243개안기산;기편마단백적상대분자질량위26 800,예측등전점위5.49,소수안기산점47.7%,친수안기산점26.8%,산성안기산점12.7%,감성안기산점12.8%;해단백유잠재적2개락단백격매Ⅱ린산화위점、3개단백격매C린산화위점화7개N-육두구선위점.결론 해안기산서렬속우용혈린지매양공능역특정단백매;여구포자균친원관계최근.통과연구,일개PM용혈린지매기인피성공발현,저위진일보탐토해기인적공능제공료조건.
Objective To identify the gene encoding lysophospholipase from the full length cDNA library of Penicllium marneffei (PM) in yeast phase and predict the structure and function of its deduced protein, to provide with theoretical evidences for further experiments. Methods The PM full-length cDNA library in yeast phase was screened to identify the gene encoding lysophospholipase with the help of NCBI on-line analytical tool. Then, by utilizing the software package of Vector NTI suite 8.0, the protein deduced by the gene was analyzed to predict its corresponding structure and functions, and its molecular cladogram was constructed. Results The gene was composed of 1014 base pairs in the length and was presumed to be the full-length gene encoding lysophospholipase by Blastx, with a complete open reading frame (ORF) comprised of 729 base pairs encoding 243 amino acids with relative molecular weight being 26 800 and the predicted isoelectric point being S.49. The deduced protein included 47.7% hydrophobic amino acids, 26.8% hydrophilic amino acids, 12.7% acid amino acids and 12.8% basic amino acids. The protein had 2 potential casein kinase Ⅱ phosphorylation site, 3 potential protein kinase C phosphorylation site and 7 N-myristoyl site. Conclusions The amino acid sequence belongs to this kind of protease with lysophospholipase-like domain. The protein is most close to Coccidioides posadasii in genetic relationship. A novel gene encoding lysophospholipase is successfully found and the work has made necessary preparations for further research on the gene's function.