Li-Li Zhang¹, Tian-Yao Bai², Xing Wan¹, Hui-Min Deng¹, Li-Ning Liang¹, Si-Chun Zheng¹, Qi-Li Feng¹ and Lin Liu¹*

¹Guangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou, 510631, China.
²Institute for Conservation and Utilization of Agro-bioresource in Dabie Mountains，School of Life Sciences, Xinyang Normal University, Xinyang, 464000, China.

 

Cholesterol is an important cellular membrane component and a precursor of biosynthesis of ecdysteroids in insects. The lack of cholesterol would retard insect development. However, insects cannot synthesize cholesterol de novo due to lack of two key enzymes in the cholesterol biosynthesis pathway, and as a result there is need for sterols uptake from food by the insects. Sterol carrier protein (SCP) is a cholesterol-binding protein responsible for cholesterol uptake and transport in insect cells and potential drug target for insecticide development. To study the function of SlSCP2 and to screen efficient and specific insect growth chemical inhibitors that target at this protein, a stably transfected CHO cell line that expresses Spodoptera litura sterol carrier protein 2 (SlSCP2) was established. Sequencing and restriction enzyme digestion analysis showed that SlSCP2 cDNAs was cloned into pcDNA5/FRT vector, which was used to transfect CHO cells, followed by drug resistance selection. Genomic PCR, RT-PCR and Western blot analysis indicated that the SlSCP2 gene integrated into the genome of the CHO cells and the mRNA and protein of SlSCP2 were correctly expressed in the transfected cells. The positive cell colonies could stably express the protein after twenty-seven (27) generations. The established transfected CHO cell line with stable expression of SlSCP2 provide a useful cell model for high throughput screening for insect growth inhibitors that target SlSCP2, as well as, for functional study of SlSCP2.

Key words: Sterol carrier protein, cholesterol, stable transfected cell line, insect growth inhibitors, high throughput screening.