EXPRESSION OF HUMAN PITUITARY ADENYLATE-CYCLASE ACTIVATING POLYPEPTIDE (PACAP) CDNA IN CHO CELLS AND CHARACTERIZATION OF THE PRODUCTS
EXPRESSION OF HUMAN PITUITARY ADENYLATE-CYCLASE ACTIVATING POLYPEPTIDE (PACAP) CDNA IN CHO CELLS AND CHARACTERIZATION OF THE PRODUCTS
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DOI：
10.1016/0014-5793(92)80020-h
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发表时间：
1992-02-17
期刊：
影响因子：
3.5
通讯作者：
FUJINO, M
中科院分区：
文献类型：
--
作者：
OKAZAKI, K;KIMURA, C;FUJINO, M
在神经内分泌的中国仓鼠卵巢细胞CHO-K1中表达人PACAP前体基因，电穿孔转染人PACAP基因的表达载体(PTS705)。建立了条件培养液中产生80 ng/ml以上免疫活性PACAP(ir-PACAP)的细胞系。对该细胞系培养上清液的反相高效液相色谱分析表明，存在两种PACAP，即PACAP38和PACAP27。同时，还发现PACAP免疫活性相关肽(ir-PRP)被分泌到培养基中。Ir-PACAPs在大鼠嗜铬细胞瘤PC12h细胞中具有诱导cAMP和突起生长等生物活性。
cDNA encoding human PACAP precursor was expressed in on-neuroendocrine Chinese hamster ovary cells, CHO-K1, The cells were transfected with expression vector (pTS705) containing the human PACAP cDNA by electroporation. A cell line which produced more than 80 ng/ml of immunoreactive PACAP (ir-PACAP) into the conditioned medium was established. RP-HPLC analysis of culture medium of this established cell line exhibited the presence of two types of PACAP, i.e. PACAP38 and PACAP27. At the same time, it was also revealed that immunoreactive PACAP-related peptide (ir-PRP) was secreted into the cultured medium. The ir-PACAPs were confirmed to have biological activities such as induction of cAMP and neurite outgrowth in rat pheochromocytoma PC12h cells.