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机构地区:[1]华中农业大学生命科学技术学院农业微生物学国家重点实验室
出 处:《实验生物学报》2003年第6期476-481,共6页Acta Biologiae Experimentalis Sinica
基 金:国家自然科学基金资助(30080013);国家863计划项目(2001AA214011);教育部重点项目资助
摘 要:S层(Slayer)是由单一的蛋白或糖蛋白组成的薄层晶状结构,它广泛存在于古细菌和真细菌细胞的最外表面,可包裹整个细胞。S层在结构化学、形态学、遗传学以及物理化学等方面具有独特的性质,使之在生物技术、分子纳米技术和仿生学等领域蕴藏着广泛的应用潜力。近年来。Cell surface layer (S-layer) is the outmost structure of bacterium cell and covers the intact cell. S-layer protein of Bacillus thuringiensis strain CTC (CTC protein) was used as carrier protein to display polyhistidine (polyHis) peptides on the cell surface. Series fusion protein genes were constructed by inserting DNA fragments encoding (6His)1, (6His)2, (6His)3, (6His)9 and (6His)15 polyHis peptides into CTC protein gene at the site of downstream of slh domain. With the help of re-combinant plasmid pBMB-CSA, which contains operon csaAB needed for S-layer protein anchoring on cell surface, fusion protein genes were expressed in crystal negative B. thuringiensis strain 4Q7. SDS-PAGE profiles demonstrated that all the fusion proteins except CTC-(6His)15 were expressed. Ni-NTA-agarose beads binding test showed that all recombinant strains could attach to agarose beads except 4Q7 (pBMB-CSA, pBMB-SH15). Cd2+ adsorption test indicated that the adsorption ability of all recombinant strains except 4Q7 (pBMB-CSA, pBMB-SH15) were higher than that of host strains. The Cd2+ adsorption quantity of the recombinant strain with strongest adsorption ability was twice higher than that of host strain.
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