Zheng Huiling, Zhu Zhenzhen, An Junhui, Yang Zhenyu, Xing Ruifang, Yan Linhui
College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.
Sheng Wu Gong Cheng Xue Bao. 2010 Dec;26(12):1704-9.
deltaFosB, a naturally occurring truncated isform of fosB gene, existed in many tissues stably and played an important role in formation and differentiation of adipocyte and osteoblast. deltaFosB may be related to the metabolism of calcium in bone and mammary gland and regulate the signal pathway of calcium transfer from bone to mammary gland. We first sub-cloned deltafosB gene of goat into the vector pET32a to construct prokaryotic expression vector pET32a-deltafosB. Then we induced for deltafosB gene expression efficiently by IPTG. Finally we immunized the adult rabbits with purified recombinant deltaFosB to prepare rabbit anti-goat deltaFosB polyclonal antibody. iELISA analysis showed the antibody with the titer of 1:51 200, and Western blotting result showed that the antibody could specifically detect the deltaFosB protein expressed in prokaryotic cell and HEK-293 cell, respectively. Further Western blotting assay showed that deltaFosB expressed in various tissues of goat in vivo.
δFosB是fosB基因天然存在的截短异构体，稳定存在于多种组织中，在脂肪细胞和成骨细胞的形成与分化中发挥重要作用。δFosB可能与骨骼和乳腺中的钙代谢相关，并调节钙从骨骼向乳腺转移的信号通路。我们首先将山羊的δfosB基因亚克隆到载体pET32a中，构建原核表达载体pET32a-δfosB。然后用异丙基-β-D-硫代半乳糖苷（IPTG）高效诱导δfosB基因表达。最后用纯化的重组δFosB免疫成年兔，制备兔抗山羊δFosB多克隆抗体。间接酶联免疫吸附测定（iELISA）分析显示该抗体效价为1:51 200，蛋白质印迹结果表明该抗体能够分别特异性检测原核细胞和人胚肾293（HEK-293）细胞中表达的δFosB蛋白。进一步的蛋白质印迹分析表明，δFosB在山羊体内的各种组织中均有表达。
