CAJ | 학술논문

目的:将L-门冬酰胺酶抗原表位区域的两个肽段作为突变对象,构建9个突变体,并对其酶活力和蛋白表达量与原酶进行比较研究.方法:根据丙氨酸扫描原理,应用重叠延伸PCR技术构建突变体,并对突变菌株进行基因测序和酶活力测定.结果:构建成功的突变菌株均具有酶活力,且蛋白表达水平不受影响.结论:首次采用基因修饰,改变L-门冬酰胺酶抗原表位区域的肽段,而不影响其酶活力,为后续研究和探讨L-门冬酰胺酶抗原表位结构与功能的关系提供了基础.
목적:장L-문동선알매항원표위구역적량개태단작위돌변대상,구건9개돌변체,병대기매활력화단백표체량여원매진행비교연구.방법:근거병안산소묘원리,응용중첩연신PCR기술구건돌변체,병대돌변균주진행기인측서화매활력측정.결과:구건성공적돌변균주균구유매활력,차단백표체수평불수영향.결론:수차채용기인수식,개변L-문동선알매항원표위구역적태단,이불영향기매활력,위후속연구화탐토L-문동선알매항원표위결구여공능적관계제공료기출.
AIM:To construct nine novel L-asparaginase mutants and study their enzyme-activity.METHODS:The mutants were constructed using overlap extension PCR according to the principle of alanine-scanning mutagenesis. The enzyme-activity was detected by Nessler's method. RESULTS:The DNA sequencing showed that the mutagenesis was consistent with the theoretical prediction. The enzyme-activity assay demonstrated that each mutant possessed enzyme activity equal to the original enzyme. CONCLUSION:Through gene modification,epitop of L-asparaginase was changed without activity loss.These results provide foundation for further study of the structure-function relationship of L-asparaginase.