Expression and characterization of recombinant rat placental prolactin-like protein C.
Expression and characterization of recombinant rat placental prolactin-like protein C.
复制标题
重组大鼠胎盘催乳素样蛋白C的表达和表征。
DOI：
10.1016/0303-7207(94)90193-7
复制
发表时间：
1994
影响因子：
4.1
通讯作者：
Shiverick,KT
中科院分区：
文献类型：
--
作者：
Conliffe,PR;Farmerie,WG;Charles,GD;Buhi,WC;Kelly,PA;Simmen,RC;Shiverick,KT
Prolactin-like protein C (PLP-C) is a member of the rat placental family of proteins which are structurally related to pituitary prolactin (PRL). In an effort to characterize the receptor specificity and biological activity of PLP-C, we used a PLP-C cDNA to express the recombinant protein in a bacterial system. The PLP-C cDNA was modified by oligonucleotide mutagenesis and ligated into a human carbonic anhydrase II (hCAII) expression vector. Following a single step affinity purification, the hCAII-PLP-C fusion protein was digested with enterokinase to release a 25 kDa protein. N-Terminal sequence analysis of the 25 kDa band demonstrated identity with PLPC. A polyclonal antiserum to the fusion protein cross reacted with seven major proteins in rat placental culture media of which two were the native forms of PLP-C. Recombinant PLP-C was not mitogenic in the Nb2 lymphoma bioassay and did not exhibit high affinity binding to rat PRL receptor. The choice of hCA-II fusion allows for rapid purification of rPLP-C which will aid in further investigation of the biological role of PLP-C.
