【作者】 高家合； 李梅云； 李天飞； 宋春满； 廖文程； 王革；

【Author】 Gao Jiahe Li Meiyun La Tianfei Song Chunman Liao Wencheng Wang Ge ( Yunnan Tobacco Research Institute, Yunnan Yuxi 653100; Yuxi Hongta Group, Yuxi 653100)

【机构】 云南省烟草科学研究所； 云南玉溪红塔集团；

【摘要】 采用液体双向分离法分离提纯苏云金杆菌Bt33、Bt53、BtHB伴孢晶体,Bt33、Bt53、BtHB分离纯度分别达99.93%、99.9%、99.8%。扫描电镜和透射电镜观察发现,3个菌株均有菱形、长椭圆形、不规则形晶体,个别有方形,少部分长椭圆形晶体为不标准椭圆形,带有“拖尾”或突起;不同菌株在晶体形状和数量组成上有所不同,Bt33菌株菱形晶体占75%,长方形与椭圆形晶体占22%-23%,Bt53菌株长椭圆形晶体占90%,菱形晶体占5%,不规则形占5%,BtHB菌株菱形晶体占30%,长椭圆形为64%-65%,方形5%;晶体大小不均一。以SDS-聚丙烯酰胺凝胶电泳分析了3个对烟草粉螟高毒力苏云金芽孢杆菌的伴孢晶体,结果发现3个菌株的电泳图谱不同,Bt33有2条清晰带,分子量分别为43kD、15kD,BtHB菌株有3条清晰的带,分子量分别为43kD、23kD、15kD,Bt53菌株有3条带,分子量分别约43kD、15kD、19kD,色淡。SDS-PAGE分析表明,一个Bt菌株有几种不同的晶体蛋白,不同菌株晶体蛋白的种类有所同亦有所不同。更多还原

【Abstract】 Parasporal crystals of 3 Bacillus thuringiensis strains were extracted by the method of liquid diphasic separation, which purity were respectively Bt33 99.93%, Bt5399.9%, BtHB 99.8% .The Mowing were found under scanning electron microscope and transmission electron microscope that all of 3 strains have rhombus, long oval, irregular form crystal, individual contain square crystal, little long oval crystal is not the standard oval which have "tail" or protuberance; Different strain has different morphology crystal.The rhombus crystal of Bt33 occupy 75%, the rectangular and oval crystal occupy 22%-23%, for Bt53 the long oval crystal occupy 90%, the rhombus 5%, irregular form 5% , but BtHB has 30% rhombus crystal, 64%-65% long oval, and 5%square.The crystal size is not uniform also.The parasporal crystal from 3 Bacillus thuringiensis strains, which were toxic highly on Ephesia elutella Hubner, were analysized by the method of SDSPAGE.The pattern of SDS-PACE electrophoresis indicated that Bt33 had two clear bands of 43kD and 15kD, BtHB had three clear bands of 43kD, 23kD and 15kD, however, Bt53 had three clear bands of 43kD, 15kD and 19kD.The conclusion can made from SDS- PACE that one Bacillus thuringiensis strain has several different kind crystal protein, the crystal protein from different isolates was same or different.更多还原