Wang Wen-Bing, Yao Bin, Xiao Qing-Li, Ji Ping, Wang Sheng-Peng, He Jia-Lu, Wu Xiang-Fu
Key Laboratory of Silkworm Biotechnology, Ministry of Agriculture, Zhenjiang 212018, China.
Sheng Wu Gong Cheng Xue Bao. 2003 Jan;19(1):112-5.
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 BmN 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.
将黑曲霉963的植酸酶基因克隆到杆状病毒转移载体中。将重组载体的DNA与Bm - BacPAK6 DNA共转染到BmN细胞中,并通过噬斑测定法筛选重组病毒。用植酸酶基因特异性探针通过斑点印迹和Southern印迹鉴定重组病毒。植酸酶基因在家蚕幼虫和蛹中表达。表达产物在蚕幼虫血淋巴中的含量为1.43 g/L,在蛹血淋巴中的含量为1.90 g/L。本文研究了杆状病毒表达系统中表达的植酸酶的酶学特性。