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[目的]分析NaCl对寡聚化的影响,以及寡聚化对Cry1Ah杀虫活性的影响。[方法]利用Superdex-75分子筛,将Cry1Ah活性片段的寡聚体和单体进行分离,通过戊二醛交联试验分析寡聚化程度,结合生物活性测定,分析NaCl对寡聚化的影响以及寡聚化对Cry1Ah毒力的影响。[结果]在纯化的组分峰Ⅰ中寡聚体占总蛋白的63.35%。NaCl对寡聚化有促进作用,而寡聚体又以多种形式存在,并且表现出不同的生物活性。寡聚体峰Ⅰ、寡聚体峰Ⅱ、单体峰Ⅲ对棉铃虫的LC50分别为47.192、3.329、3.977μg/mL。[结论]NaCl对寡聚化有促进作用,而且,寡聚体的毒力比Cry1Ah活性片段的毒力有不同程度的降低,最多下降49.6倍。推测这种体外形成的寡聚体不同于昆虫体内形成的寡聚体,引起插膜穿孔能力下降,导致毒力不同。
[Objective] The research aimed to analyze the influence of NaCl on oligomerization and the effect of oligomerization on the insecticidal activity of Cry1Ah. [Method] The oligomers and monomers of CrylAh active fragment were separated by Superdex-75 molecular sieve. The degree of oligomerization was analyzed by glutaraldehyde cross-linking test, and the effect of NaCl on oligomerization was analyzed in combination with bioactivity determination. Effect of Oligomerization on Cry1Ah Toxicity. [Results] The oligomer in the purified component peak I accounted for 63.35% of the total protein. NaCl promotes oligomerization, whereas oligomers are present in various forms and exhibit different biological activities. The LC50 of oligomer peak Ⅰ, oligomer peak Ⅱ and monomer peak Ⅲ against H. armigera were 47.192, 3.299 and 3.977 μg / mL, respectively. [Conclusion] NaCl promoted the oligomerization. Moreover, the virulence of oligomers was reduced by 49.6 folds compared with that of Cry1Ah. Speculated that the oligomers formed in vitro is different from the oligomers formed in the insect body, causing the ability of the intervertebral perforation decreased, resulting in different virulence.