CAJ | 학술논문

目的在组学水平高通量、系统地预测及筛选细粒棘球绦虫的分泌蛋白组,研究分泌蛋白及其信号肽特征,为后续诊断和疫苗相关抗原(簇)研究奠定基础. 方法利用SignalP4.1对细粒棘球绦虫全基因组蛋白序列进行信号肽预测,对含有信号肽的蛋白依次应用生物信息学软件TMHMMv2.0、Phobius、Big-PI predictor及TargetP1.1进行细粒棘球绦虫经典分泌蛋白质组筛选.随后利用LipoP1.0和TatP1.0分析分泌蛋白信号肽酶切位点类型,用SPSS19.0统计归纳信号肽和分泌蛋白的氨基酸序列特征,Blast2GO对分泌蛋白质组进行基因本体(gene ontology,GO)注释与聚类. 结果在细粒棘球绦虫14 235条全基因组序列所编码的蛋白中,共发现984条含有信号肽的蛋白序列;其中有363条属于膜结合蛋白,另有25条定位于亚细胞器;最终筛选到596条经典分泌蛋白序列.分泌蛋白质组的信号肽长度集中于15～ 31个氨基酸残基,其中疏水性氨基酸含量为62％,主要被Ⅰ型信号肽酶所识别,且酶切位点在-3及-1位相对保守;分泌蛋白氨基酸含量集中于50～ 700个氨基酸残基,小于非分泌蛋白氨基酸含量(t=3.06,P＜0.01);GO分析显示分泌蛋白主要参与新陈代谢、细胞过程、调节和发育等生物过程,并行使催化、结合、抗氧化和酶调节功能. 结论本研究预测筛选到含有596条蛋白序列的细粒棘球绦虫分泌蛋白组,可用于后续的诊断和疫苗相关抗原筛选研究.
목적 재조학수평고통량、계통지예측급사선세립극구조충적분비단백조,연구분비단백급기신호태특정,위후속진단화역묘상관항원(족)연구전정기출. 방법 이용SignalP4.1대세립극구조충전기인조단백서렬진행신호태예측,대함유신호태적단백의차응용생물신식학연건TMHMMv2.0、Phobius、Big-PI predictor급TargetP1.1진행세립극구조충경전분비단백질조사선.수후이용LipoP1.0화TatP1.0분석분비단백신호태매절위점류형,용SPSS19.0통계귀납신호태화분비단백적안기산서렬특정,Blast2GO대분비단백질조진행기인본체(gene ontology,GO)주석여취류. 결과 재세립극구조충14 235조전기인조서렬소편마적단백중,공발현984조함유신호태적단백서렬;기중유363조속우막결합단백,령유25조정위우아세포기;최종사선도596조경전분비단백서렬.분비단백질조적신호태장도집중우15～ 31개안기산잔기,기중소수성안기산함량위62％,주요피Ⅰ형신호태매소식별,차매절위점재-3급-1위상대보수;분비단백안기산함량집중우50～ 700개안기산잔기,소우비분비단백안기산함량(t=3.06,P＜0.01);GO분석현시분비단백주요삼여신진대사、세포과정、조절화발육등생물과정,병행사최화、결합、항양화화매조절공능. 결론 본연구예측사선도함유596조단백서렬적세립극구조충분비단백조,가용우후속적진단화역묘상관항원사선연구.
Objective To predict and screen the secretome of Echinococcus granulosus by using genome-wide prediction and bioinformatic approaches,and systematically analyze the characteristics of signal peptides and secreted proteins so as to provide a platform for finding diagnostic and vaccine target antigen(s).Methods The genome-wide prediction and bioinformatic methods were used in the study to profile the secretome.The signal peptides were identified from the E.granulosus whole genome sequence by using SignalP4.1 program,and the proteins containing signal sequences were analyzed by TMHMM v2.0,Phobius,Big-PI predictor and TargetP1.1 in a stepwise way to identify the classical secreted proteins.Subsequently,the enzyme digestion sites of the signal sequences identified were analyzed by LipoP1.0 and TatP1.0,and the sequence features of both the signal peptides and the secreted proteins were statistically analyzed by SPSS19.0.Finally,the ontology(GO) annotations and clustering were performed for the secretome sequences by using Blast2GO.Results A total of 984 proteins encoding sequences containing signal peptides were found in 14 235 E.granulosus genome sequences.Among them,363 sequences were membrane-binding proteins and 25 proteins were located in the subcellular organelle.Finally,a total of 596 proteins were recognized as classical secreted proteins.The signal sequences contained mostly 15 ～ 31 amino acids,among them 62％ were hydrophobic.They were recognized mainly by type Ⅰ peptide enzyme,and comparably conservative at the digestion sites-3 and-1.Most of secreted proteins identified possessed 50 ～ 700 amino acids,which was significantly less than the non-secreted ones.GO analysis showed that these secreted proteins mainly involve in metabolic,cellular,and developmental processes,and in biological regulation,playing the functions in catalytic activity,binding,antioxidant activity and enzyme regulator activity.Conclusion E.granulosus secretome containing 596 protein sequences was predicted thus to provide a database for further identification of diagnostic and vaccine target antigens.