Construction Of Transgenic Vectors Of Adipose Tissue Specific Expression Pig (Mouse) Glil

Adipose tissue is an important regulator of metabolic homeostasis and immunesystem. However, over accumulation of adipose in human body may result in obesity,diabetis and cardiovascular disease, in demestic animal’s body may decrease leanmeat percentage of carcass which is asscociated with the profit. The adiposeaccumulation is a complex dynamic process, and this process is regulated by manykinds of hormone and signal pathways. Recently Hedgehog signaling pathway plays aconserved role in inhibiting fat formation from invertebrates to vertebrates, but themolecular mechanism remains unclear. Gli1is the transcription factor which is theeffctor of Hh signal pathway to activate the target genes. But little is known aboutwhether Gli1plays a role or not when Hh inhibits adipose accumulation.In this study, we cloned the full lenth cDNA of pig Gli1through RT-PCR andRACE technique, then conducted bioinformatics and expression profile analysis. Thecoding sequences of mouse Gli1was cloned using RT-PCR technique and overlapextension PCR. The transgenic vectors of adipose tissue specific expression pig (ormouse) Gli1were construced. This work lay good foundation for generatedtransgenic pig (or mouse) with adipose tissue specific expression Gli1which can beused to investigate the mechanism of Gli1inhibition on adipose accumulation. Themain results were as follows:(1) Pig Gli1full-length cDNA sequence (3576bp) was first cloned, submitted toGenBank and gain a GenBank access number JQ246507. It was composed of12exons (including3321bp CDS region) which encoded1106amino acids. Afterthe analysis of amino acids homology among kinds of mammals, pig Gli1had thehighest homology with cow (94%), and lowest with mouse (85%). Phylogenetictree of analysis supported these resutls.(2) RT-PCR results of expression in different organs showed that pig Gli1expresshighly in pig tongue tissue, but it was not detected in heart, liver, spleen, lung,kidney, muscle, adipose and ovarian.(3) Bioinformatic analysis showed that pig Gli1is a hydrophilic protein without the transmembrane structure and signal peptide. There were five zinc finger C2H2type domains in pig Gli1.(4) The transgenic vector of adipose tissue specific expression pig Gli1wassuccessfully construced.(5) Mouse Gli1gene coding sequences were cloned，totally3336bp including with13exons and encoding1111amino acids. The transgenic vector of adipose tissuespecific expression mouse Gli1was successfully construced.