Arora Naresh, Selvapandiyan A, Agrawal Neema, Bhatnagar R K
International Centre for Genetic Engineering and Biotechnology, PO Box 10504, Aruna Asaf Ali Marg, New Delhi 110067, India.
Biochem Biophys Res Commun. 2003 Oct 10;310(1):158-62. doi: 10.1016/j.bbrc.2003.08.137.
Vegetative insecticidal protein (VIP) is a class of insecticidal proteins produced by some strains of Bacillus thuringiensis during the vegetative stage of their growth. Unlike delta-endotoxins which are produced as parasporal inclusion bodies within the cell during sporulation, VIP is secreted into the culture medium. Here we report the relocation of the expression of VIP into the mother cell compartment in a manner similar to well-characterized Cry proteins. Relocation of VIP is directed to mother cell by placing its synthesis under sporulation-dependent promoters, BtI and BtII. The insertion of cry preferred transcription termination sequence at the 3(') region and a STAB-SD sequence at the 5(') region of the gene provided stability to the vip transcript and enhanced its yield. The demonstrated expression of VIP within the cells in the form of inclusion bodies would facilitate development of a suitable formulation for the application of this class of insecticidal proteins in the field.
营养期杀虫蛋白(VIP)是苏云金芽孢杆菌某些菌株在生长的营养阶段产生的一类杀虫蛋白。与在孢子形成过程中作为细胞内伴孢晶体产生的δ-内毒素不同,VIP分泌到培养基中。在此,我们报道了VIP的表达以类似于已充分表征的Cry蛋白的方式重新定位到母细胞区室中。通过将其合成置于依赖孢子形成的启动子BtI和BtII的控制下,VIP的重新定位被导向母细胞。在基因的3'区域插入cry优先转录终止序列以及在5'区域插入一个STAB-SD序列,为vip转录本提供了稳定性并提高了其产量。以包涵体形式在细胞内证实的VIP表达将有助于开发一种适合在田间应用这类杀虫蛋白的制剂。