Li X, Liu M, Bai X, Li Y, Zhao Y, Wang S, Wang J
Jilin Ginseng Academy, Changchun University of Chinese Medicine, Changchun, Jilin 130117, China.
Pol J Vet Sci. 2019 Mar;22(1):143-150. doi: 10.24425/pjvs.2018.125613.
Osteocalcin is a major non-collagenous component of the bone extracellular matrix and is considered to be an indicative factor of osteoblast differentiation. In the present study, we detected osteocalcin expression in different antler areas and growth phases by immunohisto- chemistry. Osteocalcin was highly expressed in all areas during the mineralization period and in mesenchymal cell and chondrocyte areas during the rapid growth period. The nucleotide sequence of the osteocalcin gene in sika deer antler was determined. The open reading frame was 303 bp encoding a protein of 100 amino acids. The estimated molecular mass of osteocalcin was 10.38 kDa and the theoretical isoelectric point was 5.37. The osteocalcin gene with a 6× His-tag at the C-terminus was cloned into the pGEX-4T1 vector and expressed in Escherichia coli under optimal conditions. The recombinant soluble protein fused with GST was purified with Ni-NTA resin. The purified osteocalcin protein exhibited a significant increase in HA adhesion and promoted antler chondrocyte proliferation. Osteocalcin is an important factor in regulating the rapid growth and differentiation of deer antlers.
骨钙素是骨细胞外基质的主要非胶原蛋白成分，被认为是成骨细胞分化的指示因子。在本研究中，我们通过免疫组织化学检测了骨钙素在鹿茸不同部位和生长阶段的表达情况。在矿化期，骨钙素在所有部位均高表达；在快速生长期，骨钙素在间充质细胞和软骨细胞区域高表达。测定了梅花鹿鹿茸中骨钙素基因的核苷酸序列。其开放阅读框为303 bp，编码一个由100个氨基酸组成的蛋白质。骨钙素的估计分子量为10.38 kDa，理论等电点为5.37。将在C末端带有6×His标签的骨钙素基因克隆到pGEX-4T1载体中，并在最佳条件下在大肠杆菌中表达。与GST融合的重组可溶性蛋白用Ni-NTA树脂纯化。纯化后的骨钙素蛋白在与透明质酸（HA）结合方面显著增加，并促进鹿茸软骨细胞增殖。骨钙素是调节鹿茸快速生长和分化的重要因子。
