华中科技大学学报（医学）（英德文版）
華中科技大學學報（醫學）（英德文版）
화중과기대학학보（의학）（영덕문판）
JOURNAL OF HUAZHONG UNIVERSITY OF SCIENCE AND TECHNOLOGY(MEDICAL SCIENCE)
2012年
2期
299-302
,共4页
cannabinoid receptor 1%cannabinoid receptor 2%denaturing temperature%signal peptide%mammalian expression
Many researchers employed mammalian expression system to artificially express cannabinoid receptors,but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports.In present study,we demonstrated cannabinoid receptor protein was not able to be properly expressed with routine mammalian expression system.This inefficient expression was rescued by endowing an exogenous signal peptide ahead of cannabinoid receptor peptide.In addition,the artificially synthesized cannabinoid receptor was found to aggregate under routine sample denaturing temperatures (i.e.,≥95℃),forming a large molecular weight band when analyzed by immuno-blotting.Only denaturing temperatures ≤75℃ yielded a clear band at the predicted molecular weight.Collectively,we showed that efficient mammalian expression of cannabinoid receptors need a signal peptide sequence,and described the requirement for a low sample denaturing temperature in immuno-blot analysis.These findings provide very useful information for efficient mammalian expression and immuno-blotting of mcmbrane receptors.
