Biological characteristics of KLF8 and its effect on intramuscular preadipocyte proliferation in goats
This experiment aimed to clone goat KLF8 gene sequence,elucidate its tissue expression characteristics,and reveal the potential effect of KLF8 on goat intramuscular preadipocytes proliferation through overexpression technique.The promoter and CDS region sequence of goat KLF8 gene was cloned by RT-PCR technique,along with bioinformatics analysis of sequence.pcDNA3.1-KLF8 overexpression vector was constructed by gene recombination method.CCK-8,EdU and real-time quantitative PCR(qPCR)were used to clarify the expression pattern of KLF8 in the proliferation of goat intramuscular preadipo-cytes and its effect on proliferation.The results showed that the obtained goat KLF8 gene promoter sequence was 2 001bp in length,containing two potential core promoters,multiple TATA-boxes and GATA-boxes,and 1 062 bp of CDS sequence contai-ning typical zinc-finger domain of KLF family and transcriptional repressor/activation motifs.Tissue expression profile assay found that KLF8 was expressed at a high level in spleen tissue and significantly higher than any other tissues(P<0.01),at a medium level in kidney and lung tissue,at a low level in longissimus dorsi muscle and biceps femoris muscle.Overexpression of goat KLF8 inhibited the proliferation of intramuscular preadipocytes,and this regulation may be achieved mainly through up-reg-ulating the expression of CCND1.The results provided important data support to clarify the biological characteristics of goat KLF8 gene and its effect mechanism on the proliferation of goat intramuscular preadipocytes.
NETL
NSTL
万方数据
维普
