非同位素EGF-反义mRNA探针的制备及其与组织切片的原位杂交

The Preparation of Nonradioactive EGF-antisense mRNAProbes and in Situ Hybridization with Tissue Section

本实验将含大鼠ＥＧＦ基因的ｐＧＥＭ４质粒扩增后制备出ＤＮＡ模板，体外转录合成非同位素反义ｍＲＮＡ探针，并与大鼠领下腺组织切片进行原位杂交。结果显示颗粒曲管细胞内杂交信号清晰可见，表明非同位素探针与同位素探针相比不仅具有敏感性和特异性，而且安全、简便、可靠。此外本实验对建立可靠易行的原位杂交方法进行了摸索。

Abstract: In this paper the DNA template was prepared after the pGEM4 plasmid (containg rat EGF gene )was amplificated, the nonradioactive antisense mRNA probe transcripted andsynthesized in vitro. Using this probe the situ hybridization was carried out on rat submandibular section. The results showed that hybridized singnal in granular duct cell is clear. It indicatedthat nonradioactive probe not only have sensitivity and specificity, but safe,simple and reliable.In adition. to set up a reliable technic of in situ hybridization have been studied.Kdy Words: probeEGF (epidermol growth factor ); in situ hybridization