The Study Of Proteolipid Protein 2 Gene In Liaoning Cashmere Goat

Liaoning cashmere goat is the precious genetic resources of China which is banned outflow. Cashmere production ranks first in the country. In previous work, we constructed a skin cDNA library and isolated a full-length cDNA clone using Primer walking method termed proteolipid protein 2. In order to explore the function of PLP2, We firstly analyzed the cDNA and amino acid sequences by biological informatics analysis(ORF predicting, hydrophobicity analysis, transmembrane domain predicting, signal peptideand, phosphorylation site and forecasted secondary structure,evolutionary tree, multiple sequence alignment, structural domain). Otherwise, RT–PCR was used to analyze the expression of PLP2 gene in all internal organs. Real-time-PCR was used to detect the contents of PLP2 gene expression in the primary and secondary hair follicles during different growth periods. And the PLP2 gene was detected specificity expression sites by In situ hybirdization. Finally, we using qPCR to detect the expression of PLP2 gene which were infected by inhibitor-Noggin gene lentivirus.The results were as follow: firstly, The results of PLP2 showed it could encode the protein including 152 amino acids. Secondary structure had the Alpha helix(46.05%), extended strand(12.5%) and random coil(41.45%). Function site analysis showed that PLP2 had a MARVEL protein domain, four transmembrane domains and no signal peptide. PLP2 contain three Ser, two Tyr and one Thr could be the potential protein kinase phosphorylation site. RT-PCR result showed that PLP2 was expressed in skin, heart, liver, kidney, lung, spleen tissues, and the expression quantity were closed. Real-time PCR results showed that the expression of PLP2 in secondary hair follicles was 1.02 times(0.01<P<0.05) of the primary follicles during anagen; the expression of PLP2 in the second hair follicles was 1.16 times(0.01<P<0.05) of the primary follicles during catagen; In situ hybirdization showed that PLP2 was expressed in the Inner Root Sheath(IRS) but no expression in the hair shaft(Cuticle, cortex and medullary layer), Outer Root Sheath, or Follicular Papilla(FP). Finally, using qPCR to detect the expression of PLP2 gene which were infected by inhibitor-Noggin gene lentivirus, we found the liquid viral infection group was 0.64 times than the NC group, and the uninfected cell control group was 1.15 times than the NC group.By the above results, we can draw the following conclusions:(1)PLP2 was expressed in skin, heart, liver, kidney, lung, spleen tissues;(2)The expression of PLP2 in second hair follicles was higher than the primary follicles during anagen and catagen, which could be inferred that PLP2 had a potentially important role in regulating cashmere-fiber fineness;(3) PLP2 gene is expressed in the inner root sheath, which speculate PLP2 is associated with the cashmere falling off;(4)The expression of PLP2 gene being infected was decreased, which might demonstrate PLP2 genes, along with other inhibition pathway gene, such as noggin, affected the BMP pathway and the growth of hair follicle.