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《Acta Biologiae Experimentalis Sinica》 1996-04
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CLONING AND IDENTIFICATION OF RECOMBINANT cDNA TO A RABBIT OVIDUCTIN "DPF-1"

LIU Chuan Ju SHEN Hong GU Zheng LU Ji Ning CHENG Guo Xiang TSO Jia Ke (Shanghai Institute of Cell Biology, Chinese Academy of Sciences, Shanghai, 200031)  
A recombinant cDNA library to po-lyA+RNA isolated from rabbit oviduct epithelial cells was constructed, and screened with a polyclonal antibody against DPF-1 (64kDa). 4 immunopositive plaques (DPF-1.1, DPF-1.2, DPF-1.3 and DPF-1.4) were purified. The polyclonal antibodies were epitope-selected respectively against the fused proteins produced by these positive recombinant plaques. Identification of recombinant clones by epitope selection revealed that the epitope -selected antibodies from DPF-1.1, DPF-1.2 and DPF-1. 3 could recognise not only DPF-1, but 44 kDa protein also (Fig. 2). By using EcoRI-Notl digestion method, the insert cDNA fragment size of these three recombinants was revealed to be 0.8kb, 1.2kb and 1.2kb respectively (Fig. 3). These cDNA fragments were then isolated and subcloned into pBiuescriptKS, and recombinant plasmids (pDPF-1.1, pDPF-1.2 and pDPF-1.3) were con- structed (Fig.4). Dot blot hybridization with a 32p-labeled 1.2Kb-insert of cDNA from pDPF-1.3 indicated that these recombinant plasmids could cross-hybridized (Fig. 5), further indicating that they all possessed a common nucleic acid sequence. Dot and Northern blotting analysis of total RNA prepared from eight different tissues (skeleton muscle, heart, kidney, oviduct, liver, spleen, lung and small intestine) showed that the gene encoding DPF-1 was expressed specifically in the oviduct tissue (Fig. 6, Fig. 7).
【Fund】: 国家自然科学基金~~
【CateGory Index】: Q785
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