title: The Pacific White Shrimp β-actin Promoter: Functional Properties and the Potential Application for Transduction System Using Recombinant Baculovirus.
abstract: A newly isolated Pacific white shrimp (Litopenaeus vannamei) beta-actin promoter SbaP and its derivative compact construct SbaP (ENX) have recently been demonstrated to promote ectopic gene expression in vitro and in vivo. To further explore the potential transduction application, this newly isolated shrimp promoter SbaP was comparatively tested with cytomegalovirus (CMV), simian virus 40 (SV40), polyhedrin (Polh), and white spot syndrome virus immediate early gene 1 (WSSV ie1) four constitutive promoters and a beta-actin promoter (TbaP) from tilapia fish to characterize its promoting function in eight different cell lines. Luciferase quantitation assays revealed that SbaP can drive luciferase gene expression in all eight cell lines including sf21 (insect), PAC2 (zebrafish), EPC (carp), CHSE-214 (chinook salmon), GSTEF (green sea turtle), MS-1 (monk seal), 293T (human), and HeLa (human), but at different levels. Comparative analysis revealed that the promoting activity of SbaP was lower (≤10-fold) than CMV but higher (2-20 folds) than Polh in most of these cell lines tested. Whereas, SbaP mediated luciferase expression in sf21 cells was over 20-fold higher than CMV, SV40, Polh, and TbaP promoter. Compared to the SbaP, SbaP (ENX), which was constructed on the basis of SbaP by deletion of two "negative" regulatory elements, exhibited no significant change of promoting activity in EPC and PAC2 cells, but a 5 and 16 % lower promoting effect in 293T and HeLa cells, respectively. Additionally, a recombinant baculovirus was constructed under the control of SbaP (ENX), and efficient promoter activity of newly generated baculoviral vector was detected both in vitro of infected sf21 cells and in vivo of injected indicator shrimp. These results warrant the potential application of SbaP, particularly SbaP (ENX) in ectopic gene expression in future.
authors.affiliation: Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China.|University of Chinese Academy of Sciences, Beijing, 100049, China.|Department of Public Health Sciences, University of Hawaii at Manoa, Honolulu, HI, 96822, USA.
authors.forename: Yingli
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China. jhxiang@qdio.ac.cn.
authors.forename: Jianhai
authors.initials: J
authors.lastname: Xiang
authors.affiliation: Department of Public Health Sciences, University of Hawaii at Manoa, Honolulu, HI, 96822, USA.|College of Bioengineering, Henan University of Technology, Zhengzhou, 450001, China.
authors.forename: Guangzhou
authors.initials: G
authors.lastname: Zhou
authors.affiliation: Department of Human Nutrition, Food and Animal Sciences, University of Hawaii at Manoa, Honolulu, HI, 96822, USA.
authors.forename: Tetsuzan Benny
authors.initials: TB
authors.lastname: Ron
authors.affiliation: Department of Public Health Sciences, University of Hawaii at Manoa, Honolulu, HI, 96822, USA.
authors.forename: Hsin-I
authors.initials: HI
authors.lastname: Tong
authors.affiliation: Department of Public Health Sciences, University of Hawaii at Manoa, Honolulu, HI, 96822, USA.
authors.forename: Wen
authors.initials: W
authors.lastname: Kang
authors.affiliation: Department of Public Health Sciences, University of Hawaii at Manoa, Honolulu, HI, 96822, USA.
authors.forename: Si
authors.initials: S
authors.lastname: Sun
authors.affiliation: Department of Public Health Sciences, University of Hawaii at Manoa, Honolulu, HI, 96822, USA. yuanan@hawaii.edu.
authors.forename: Yuanan
authors.initials: Y
authors.lastname: Lu
mesh_terms: D000199:Actins / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005818:Genetic Engineering / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D033561:Penaeidae / Q000166:cytology / Q000235:genetics; D011401:Promoter Regions, Genetic; D012474:Salmon / Q000235:genetics; D013539:Simian virus 40 / Q000235:genetics; D018411:Spodoptera / Q000166:cytology / Q000235:genetics; D017210:Tilapia / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods*; D014426:Turtles / Q000235:genetics; D046848:White spot syndrome virus 1 / Q000235:genetics; D015027:Zebrafish / Q000235:genetics
chemical_list: D000199:Actins; D008156:Luciferases
keywords: Promoter activity; Recombinant baculovirus; SbaP; Shrimp beta-actin promoter; The Pacific white shrimp