Premium
Cloning, expression pattern analysis, and subcellular localization of Capra hircus SCD1 gene with production of transgenic mice
Author(s) -
Zuo Qisheng,
Jin Kai,
Song Jiuzhou,
Zhang Yani,
Li Bichun
Publication year - 2018
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.26386
Subject(s) - microbiology and biotechnology , capra hircus , complementary dna , gene , biology , transfection , transgene , gene expression , cloning (programming) , subcellular localization , genetics , zoology , computer science , programming language
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 F 1 mice. The expression of exogenous gene in the F 1 generation was detected in both DNA and protein. The results showed that the coding sequence (CDS) fragments of C. 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 F 1 mice generation.