Preliminary Study On Structure And Function Of Fbxo8 Gene In Swine

Pig genetic improvement are always been the attention highlight of the pig industry scientific and technological researchers.The traditional breeding methods in pig improvement have achieved remarkable successes.Along with the development and constant breakthroughs of life science and technology,especially molecular biology,the molecular breeding will become more important and effective.Economic traits are controlled by complex genetic factors.Protein is the carrier and function executor of life activities.Ubiquitin-proteasome pathway is an important protein degradation system,which is involved in many important cellular processes through degradation of many specific proteins such as cell cycle or transcription factors.The most important feature of Ubiquitin-proteasome pathway is the substrate selectivity and diversity,and this function is mainly decided directly by the ubiquitin-proteasome - E3 s ligase protein F-box.In this study,the porcine FBXO8 gene was cloned and functionally analyzed by bioinformatics and molecular biology methods, the main results are listed as follows:1.Use of the information in the network database,through bioinformatics tools, conducted the systematic bioinformatics research and electronic cloned the swine FBXO8 gene coding sequence.2.With the swine EST information database,separated and genetic acquired the swine FBXO8 sequence.3.Use RT-PCR method,the swine FBXO8 gene tissues' distribution results showed that the detection of this gene in adult Wuzhishan pig's 16 tissues there are eight tissues expressed on different levels.4.Use pEGFP-N1 vector we construct human and pig FBXO8 subcellular localization vectors.Expressed in the human lung adenocarcinoma cell line(A549),pig kidneys cells(PK15),respectively,for the people,swine's subcellular localization studies. From the human FBXO8 subcellular localization result,observed that FBXO8 mainly distributed in the cytoplasm.5.Constructed pig-FBXO8 pcDNA 3.1-expression vector,in the pig kidneys cells (PK15),accomplished a FBXO8 over-expression.The expression in the cells after 32 hours,there was no obvious morphological changes,use RT-PCR acquire comparison sample cell DNA.6.Successfully constructed the swine FBXO8 prokaryotic expression vector,laid the foundation for the pig FBXO8 protein functional and follow-up studies.