Moroto T, Kamada A
Department of Biochemistry, Osaka Dental University, Japan.
J Osaka Dent Univ. 1998 Oct;32(2):37-42.
We investigated the characteristics and mRNA expression of hepatocyte growth factor (HGF) in the adult female monkey submandibular gland using Western blot analysis and the reverse-transcriptase PCR (RT-PCR) technique, and observed HGF mRNA localization using the in situ RT-PCR technique. HGF was extracted with PBS containing protease inhibitors, and purified partially by heparin affinity chromatography. With Western blot analysis, the HGF fraction showed an immunopositive protein band of 69-kDa corresponding to the alpha-subunit of HGF. The gene expression for HGF was revealed in the monkey submandibular gland using RT-PCR, and the PCR products showed high homology to cDNA for human HGF. Furthermore, the mRNA signal for HGF was localized in the striated and excretory ductal cells. These results unequivocally confirmed both the synthesis and existence of HGF in monkey submandibular glands.
我们采用蛋白质免疫印迹分析和逆转录聚合酶链反应(RT-PCR)技术,研究成年雌性猴下颌下腺中肝细胞生长因子(HGF)的特征和mRNA表达,并运用原位RT-PCR技术观察HGF mRNA的定位。用含蛋白酶抑制剂的磷酸盐缓冲液(PBS)提取HGF,并通过肝素亲和层析进行部分纯化。通过蛋白质免疫印迹分析,HGF组分显示出一条对应于HGFα亚基的69-kDa免疫阳性蛋白条带。利用RT-PCR在猴下颌下腺中检测到了HGF的基因表达,PCR产物与人HGF的cDNA具有高度同源性。此外,HGF的mRNA信号定位于纹状管和排泄管细胞中。这些结果明确证实了猴下颌下腺中HGF的合成与存在。