Promoter detection and transcriptional analysis of the spinosad biosynthetic gene cluster.
- Author:
Xiaozhou FENG
1
;
Weishan WANG
;
Xiaohui REN
;
Xinli LIU
;
Xiangzhao MAO
;
Keqian YANG
Author Information
1. Shandong Provincal Key Laboratory of Microbial Engineering, School of Food and Bioengineering, Jinan 250353, Shandong, China.
- Publication Type:Journal Article
- MeSH:
Drug Combinations;
Insecticides;
metabolism;
Macrolides;
metabolism;
Multigene Family;
Plasmids;
Promoter Regions, Genetic;
Real-Time Polymerase Chain Reaction;
Saccharopolyspora;
genetics;
metabolism
- From:
Chinese Journal of Biotechnology
2013;29(7):914-926
- CountryChina
- Language:Chinese
-
Abstract:
Spinosad represents a new class of insecticides produced by Saccharopolyspora spinosa. To understand the transcription of the spinosad biosynthetic gene cluster, two promoter detection plasmids based on different reporter genes were constructed and used to detect 9 promoters in the spinosad biosynthetic gene cluster. In addition, the temporal transcriptional profiles of the corresponding genes controlled by the 9 promoters, together with 4 genes outside of the spinosad cluster but are required for the synthesis of sugars in spinosad, were examined by real-time PCR. The results indicate that the 9 spinosad biosynthetic genes were highly expressed at the stationary phase, which coincides with the accumulation of spinosad in the fermentation broth. Of particular note is that the transcription of the 4 sugar synthetic genes showed higher level at the exponential phase, suggesting the expression of sugar synthetic genes is not correlated with the spinosad synthetic genes. The data suggest that spinosad biosynthesis could be improved by engineering the expression pattern of the sugar synthetic genes that lie outside the spinosad gene cluster.
