植酸酶基因在家蚕-杆状病毒表达系统中的表达及其酶学特性  

Expression of Phytase Gene in Bombyx Mori

作　　者：王文兵[1] 姚斌[2] 肖庆利[1] 季平[1] 汪生鹏[1] 何家禄[1] 吴祥甫 

机构地区：[1]中国农业科学院蚕业研究所农业部家蚕生物技术重点开放实验室,镇江212018 [2]中国农业科学院饲料研究所,北京100081 [3]上海生物化学与细胞研究所,上海200031

出　　处：《生物工程学报》2003年第1期112-115,共4页Chinese Journal of Biotechnology

基　　金：国家"8 63"高技术项目基金资助 (No.10 2 11 0 2 0 6)~~

摘　　要：将从黑曲霉菌株Aspergillusniger 96 3克隆并经改造后的植酸酶基因在昆虫 -杆状病毒表达系统中表达 ,SDS PAGE电泳检测蚕体和蛹的表达量分别达到 1.4 3g L血淋巴液和 1.90g L血淋巴液。酶活性测定结果表明 ,在蚕体和蛹的表达活性分别为 4 .6 7× 10 8u L血淋巴液和 5 .99× 10 8u L血淋巴液。该酶活性的最适温度范围为 5 0～6 0℃ ,最适pH值为 5 .5～ 5 .0和 2 .5。研究表明杆状病毒系统表达的植酸酶具有耐酸性和抗高温的特性 ,可以用于生产饲用植酸酶。Phytase gene of Aspergillus niger 963 was cloned into baculovirus transfer vector. DNA of the recombinant vector was co transfected with Bm BacPAK6 DNA into Bm N cells, and recombinant virus was selected by plaque assays. The recombinant virus was identified by Dot blot and Southern blot with the specific probe for phytase gene. Phytase gene was expressed in silkworm larvae and pupae. The expression product was 1.43 g/L haemolymph for silkworm larvae and 1.90 g/L haemolymph for pupae, respectively. The enzymic characteristicses of phytase expressed in baculovirus expression system were studied in this paper.

关 键 词：植酸酶基因 家蚕-杆状病毒表达系统 基因表达 酶学特性 

分 类 号：Q789[生物学—分子生物学] S816.79[农业科学—饲料科学]