Cloning, Expression Pattern Analysis and Subcellular Localization of Capra hircus SCD1 gene with production of Transgenic Mice

This study aimed to clone the Stearoyl‐CoA desaturase 1 (SCD1) gene derived from Xuhuai goat (Capra hircus), and analyze the sub‐cellular localization in cells and tissues. The cDNA was cloned by reverse transcription polymerase chain reaction (RT‐PCR). pEGFP‐SCD1 vector was constructed to detect sub‐cellular localization and tissue distribution. pEGFP‐SCD1 was transfected into NIH‐3T3 cells using polyethylene imine (PEI) and observed under fluorescence inversion microscope system 48 h after transfection. The expression level of SCD1 was detected by RT‐PCR. Testicular injection was used to produce transgenic mice with goat SCD1 gene. DNA and protein were extracted from the tail tissue of F1 mice. The expression of exogenous gene in the F1 generation was detected in both DNA and protein. The results showed that the coding sequence (CDS) fragments of Capra hircus SCD1 gene was 1074 bp and encodes 360 amino acids. RT‐PCR results showed that SCD1 could be expressed successfully in NIH‐3T3 cells in vitro. Sub‐cellular localization analysis showed that pEGFP‐SCD1 fusion protein located in the cytoplasm. It can be concluded that transgenic mice with goat SCD1 expressed in sperm and tail tissue was successfully produced in the F1 mice generation. This article is protected by copyright. All rights reserved

Source: Journal of Cellular Biochemistry - September 15, 2017 Category: Biochemistry Authors: Qisheng Zuo, Kai Jin, Jiuzhou Song, Yani Zhang, Bichun Li Tags: Article Source Type: research