Cloning, Expression And Expression Pattern Of Zfp36 Gene In Goat

Zinc finger protein gene zfp36 was a transcription factor with finger shape structure domain, which played an important role in regulating gene expression, cell differentiation, embryonic development, enhance plant resistance etc. It was related to the syndrome of the arthritis, autoimmune disease, hyperosteogeny in human and mice. While there were no report about goat zfp36 gene, in this study, the structure and functional areas of goat zfp36 gene were analysised, the mutation status and population specificity were augmented, the gene expression differences were studied. This paper aimed to provide the theory basis for later eliminates inflammation, perfect the inflammation mechanism.The coding sequence of Goat zfp36 gene was cloned by RT-PCR method in this study. Bioinformatics were anlysised by on line tools and softwares, such as Bioedit、BLAST、SMART、ProtParam、SCOPMA. The results showed that the length of goat zfp36 coding sequence was 978bp, coding 325 amino acids, and nucleotide sequences consistency in human, pig, ox, sheep and rats were 86%, 89%, 98%, 99%, 82% through comparing homologous sequences. Polymorphism was detected by PCR-SSCP in order to discuss the genotype regularity of distrbution in Jining grey goat, Leizhou native goat, Nanjiang Mongolian Grazelle, Liaoning Cashmere goat. And there was no SNP in exon , but a locu with A/T base displacement in intron, and the genotypes had significant difference among four goat breeds.The genetic encoding the restructuring of the zfp36 nuclear expression vector pet30-a-zfp36 was constructed, optimizing express condition, and goat zfp36 protein expressed was detected by SDS-PAGE mathod. Moreover, real-time quantitative polymerase chain reaction (RT-PCR) assays in 8 tissues of goat including lung heart, liver, spleen, lung, kidney, brain, gut, stomach showed that zfp36 was ubiquitous in all tissues examined,with the highest level in kidney and liver and the lowest in brain and heart.The results indicated that the structure and functional areas of zfp36 in species were relatively conservative, especially in exon of goat, and frequency distribution of mutation was specificity between breeds in population genetics.The size of protein expressed was 33KD, consistent as prediction, and the protein existed in soluble forms. Tissue expression profile showed that he highest level of zfp36 was expressed in kidney and liver and the lowest in brain and heart, and there were some tissue specific.