多能硫杆菌1,5-二磷酸核酮糖羧化酶/加氧酶基因的启动子克隆和表达  

FUNCONAL EXPRESSION AND SIGNAL TRANSDUCTION CROSS THE MEMBRANE OF OPIOID RECEPTER LIKE 1 RECEPTOR ORL1 IN NEUROBLSTOMA SK N SH CELLS

作　　者：刘振盈[1] 颜望明[1] 徐海岩 郑雷 罗小平 

机构地区：[1]山东大学微生物技术国家重点实验室

出　　处：《山东大学学报（自然科学版）》1997年第2期225-230,共6页Journal of Shandong University(Natural Science Edition)

基　　金：国家及山东省自然科学基金

摘　　要：将３．４ｋｂ的多能硫杆菌１，５－二磷酸核酮糖羧化酶／加氧酶基因（ｒｂｃＬ－ｒｂｃＳ）片段亚克隆到启动子探测质粒载体ｐＫＬ６的ＨｉｎｄⅢ位点，该基因片段能够启动ｐＫＬ６的ｌａｃ基因的表达，说明３．４ｋｂ的ｒｂｃＬ－ｒｂｃＳ基因带有自己的启动子．A new type of opioid receptor ORL1 and its newly identified endogenous agonist nociceptin have been implied to play an important role in pain modulation.Detected through Northem blot analysis,native expression of ORL1 receptor transcripts in human neuroblastoma SK N SH cells.Treatment of the cells with nociceptin inhibited forskolin induced intracellular accumulation of cyclic AMP,showing negative coupling of ORL1 receptor to adenylyl cyclase in SK N SH cells.IC50 of nociceptin was 60±15nmol/L,and maximal inhibition was about 70%.Treatment with DPDPE,a specific agonist of delta opioid receptor(DOR),or DAGO,a specific agonist of Mu opioid receptor (MOR) also produced inhibition of cAMP production to different extents.Further experiments found that naloxone,a nonselective antagonist of opioid receptors could block the effect of nociceptin and DPDPE.However,natrindole the specific antagonist of DOR,could only reverse the inhibition of DPDPE,but had little effect on the inhibition of nociceptin.The cellular responsiveness to nociceptin was significantly reduced after a 20 min prechallenge of the cells with 5μmol/L nociceptin ,indicative of acute desensitization of ORL1 receptor.These results suggest that ORL1 expresses in SK N SH cells with similar functions in signal transduction across the membrane as other subtypes of opioid receptor.The existence of DOR,MOR and ORL1 in SK N SH cells make it possible that SK N SH cells could be served as a good model system for studying ORL1 signal transduction and its interaction with other subtypes of opioid receptor.

关 键 词：多能硫杆菌 RUBISCO基因 无性系 启动子 

分 类 号：Q939.103[生物学—微生物学]