CAJ | 학술논문

为了解亲环素基因(cyp1.其编码亲和蛋白)在尖孢镰刀菌古巴专化型生理小种1号和生理小种4号之间及与其他镰刀菌之间的差异,分析cyp1基因与两生理小种之间的寄主选择性差异的关系,通过比对现有镰刀菌属的亲环素基因序列,设计并合成引物.采用PCR和RT-PCR方法扩增了尖孢镰刀菌古巴专化型生理小种1号和生理小种4号2个生理小种的cypl基因,并测序进行比较分析,同时对编码蛋白进行了氨基酸序列比对和功能分析.结果表明,2个生理小种cypl基因全长均为1066 bp.开放阅读框均为675 bp,编码224个氨基酸,基因序列间差异性为0.8%,编码蛋白间仅存在1个氨基酸的差异;两生理小种的cypl基因序列与镰刀菌属其他专化型或不同种间存在差异性,与不同属之间其他种间的差异最大达50%.比对蛋白序列发现仅存在0.85%的差异,这进一步证明亲和蛋白在真菌中较保守.从cyp1基因序列及其编码蛋白的氨基酸序列在两生理小种之间的差异性分析结果推测,2个生理小种寄主选择性差异与cyp1基因并无明显对应关系,这为进一步研究cypl基因功能奠定了基础.
위료해친배소기인(cyp1.기편마친화단백)재첨포렴도균고파전화형생리소충1호화생리소충4호지간급여기타렴도균지간적차이,분석cyp1기인여량생리소충지간적기주선택성차이적관계,통과비대현유렴도균속적친배소기인서렬,설계병합성인물.채용PCR화RT-PCR방법확증료첨포렴도균고파전화형생리소충1호화생리소충4호2개생리소충적cypl기인,병측서진행비교분석,동시대편마단백진행료안기산서렬비대화공능분석.결과표명,2개생리소충cypl기인전장균위1066 bp.개방열독광균위675 bp,편마224개안기산,기인서렬간차이성위0.8%,편마단백간부존재1개안기산적차이;량생리소충적cypl기인서렬여렴도균속기타전화형혹불동충간존재차이성,여불동속지간기타충간적차이최대체50%.비대단백서렬발현부존재0.85%적차이,저진일보증명친화단백재진균중교보수.종cyp1기인서렬급기편마단백적안기산서렬재량생리소충지간적차이성분석결과추측,2개생리소충기주선택성차이여cyp1기인병무명현대응관계,저위진일보연구cypl기인공능전정료기출.
In order to know the differences of cyp1 gene between Fusarium oxysporum f sp. cubense (Foc)race 1 and race 4 and other Fusarium., and analyze whether the host selective difference between the races 1 and 4 were related with Cyclophilin A (cypl) gene, the cyp1 genes of races 1 and 4 were amplified byPolymerase Chain Reaction and Reverse Transcription Polymerase Chain Reaction and sequenced, and the gene sequences and the encoded protein sequences were aligned by using the DNAStar software. The results revealed that the full length of cyp1 from Foc races 1 and 4 were both 1066bp, with their openreading frames being 675 bp, encoding 224 anuno acids. The sequences of the cyp1 gene of Foc race 1 showed 99.2% similarity to that of Foc race 4 and there was only one arruno acid different in the sequences. The cyp1 gene sequences of these two races were different from those of other specialized forms or species of Fusarium or from those of other species of different genera (the highest being 50.9% by alignment), and the protein blast result showed only 0.85% difference. This proved that the cyp1 protein was conserved in fungi,, It was inferred from the differences of the cyp1 gene sequence and protein sequences from FOC1 and FOC4 isolates that there was no obvious relation between the cyp1 gene and the host selective preference of FOC1 and FOC4. These results would help to know the function of the cypl gene from FOC1 and FOC4 in the future researches.