Functional Analysis of 5′ Upstream Control Region of Mouse Lrp5 Gene

Lv Ping, GONG Yao-Qin *, LI Jiang-Xia, ZHOU Hai-Bin, CHEN Bing-Xi

Chinese Journal of Biochemistry and Molecular Biology ›› 2005, Vol. 21 ›› Issue (05) : 616-621.

PDF(362 KB)
PDF(362 KB)
Chinese Journal of Biochemistry and Molecular Biology ›› 2005, Vol. 21 ›› Issue (05) : 616-621.
Article

Functional Analysis of 5′ Upstream Control Region of Mouse Lrp5 Gene

  • Lv Ping, GONG Yao-Qin *, LI Jiang-Xia, ZHOU Hai-Bin, CHEN Bing-Xi
Author information +
History +

Abstract

To identify the functional elements of 5′ upstream control region of mouse LDL receptor related protein 5 (LRP5) gene, the upstream region 3 041 bp (-2 909 bp ~ +132 bp) from the translation initiation codon of mouse Lrp5 gene was amplified by PCR. The PCR product was cloned into the luciferase reporter vector pGL3-basic, and the recombinant plasmid pGL3-2 909 was constructed. The different upstream regions were amplified using plasmid pGL3-2 909 as a template, thereafter inserted into the pGL3-103 vector which harboredLrp5 gene basal promoter and luciferase reporter gene. Twelve luciferase expression vectors containing 5′ upstream regions of mouseLrp5 gene were constructed, including pGL3-267, pGL3-513, pGL3-535,pGL3-560,pGL3-575,pGL3-623,pGL3-645,pGL3-719,pGL3-770,pGL3- 1 032 ,pGL3- 1 330 and pGL3-1 619 individually. All expression vectors were transfected into COS-7, respectively. Relative luciferase activities in each cell lysate were measured after 48 hours. The relative luciferase activity of pGL3-575 (-2 909 bp ~ -2 334 bp) was 20% of pGL3-513 (-2 909 bp~-2 396 bp), the activity of pGL3-535 (-2 909 bp ~ -2 374 bp) was 44% of the pGL3-513 and pGL3-575 was 48% of pGL3-560 (-2 909 bp~ -2 349 bp). The results suggest that the negative control elements existed in the 22 bp region between the -2 396 bp and -2 374 bp, as well as the 15 bp region between the -2 349 bp and -2 334 bp. IK2, LYF1 and MZF1 consensus sequences were identified using Mat Inspector V2.2 soft ware.

Key words

LDL receptor related protein 5 / mouse / Lrp5 gene / control region / luciferase activity

Cite this article

Download Citations
Lv Ping, GONG Yao-Qin *, LI Jiang-Xia, ZHOU Hai-Bin, CHEN Bing-Xi. Functional Analysis of 5′ Upstream Control Region of Mouse Lrp5 Gene[J]. Chinese Journal of Biochemistry and Molecular Biology, 2005, 21(05): 616-621

References

1 Tolwinski NS,Wehrli M,Rives A,Erdeniz N,DiNardo S,Wieschaus E.Wg/Wnt signal can be transmitted through arrowLrp5,6and Axin independently of Zw3/Gsk3beta activity.Dev Cell,2003,4(3):407~418 2 Gong Y,Vikkula M,Boon L,LiuJ,Beighton P,Ramesar R,Peltonen L,Somer H,Hirose T,Dallapiccola B,De Paepe A,Swoboda W,Zabel B,Superti-Furga A,Steinmann B,Brunner HG,Jans A,Boles R G,Adkins W,van den Boogaard MJ,Olsen B R,Warman ML.Osteoporosis-pseudoglioma syndrome,a disorder affecting skeletal strength and vision,is assignedto chromosome region11q12-13.AmJ HumGenet,1996,59:146~151 3 Gong Y,Slee RB,Fukai N,Rawadi G,Roman-Roman S,Reginato A M,Wang H,Cundy T,Glorieux F H,Lev D,Zacharin M,Oexle K,Marcelino J,Suwairi W,Heeger S,Sabatakos G,Apte S,Adkins W N,Allgrove J,Arslan-Kirchner M,Batch J A,Beighton P,Black G C,Boles R G,Boon L M,Borrone C,Brunner H G,Carle G F,Dallapiccola B,De Paepe A,Floege B,Halfhide M L,Hall B,HennekamR C,Hirose T,Jans A,Juppner H,Kim C A,Keppler-Noreuil K,Kohlschuetter A,LaCombe D,Lambert M,Lemyre E,Letteboer T,Peltonen L,Ramesar R S,Romanengo M,Somer H,Steichen-Gersdorf E,Steinmann B,Sullivan B,Superti-Furga A,Swoboda W,van den Boogaard MJ,Van Hul W,Vikkula M,Votruba M,Zabel B,Garcia T,Baron R,Olsen B R,Warman M L.Osteoporosis-pseudoglioma syndrome collaborative group.LDLreceptor-related protein5(Lrp5)affects bone accrual and eye development.Cell,2001,107(4):513~523 4 Little R D,Carulli J P,Del Mastro R G,Dupuis J,Osborne M,Folz C,Manning S P,Swain P M,Zhao S C,Eustace B,Lappe M M,Spitzer L,Zweier S,Braunschweiger K,Benchekroun Y,Hu X,Adair R,Chee L,FitzGerald MG,Tulig C,Caruso A,Tzellas N,Bawa A,Franklin B,McGuire S,Nogues X,Gong G,Allen KM,Anisowicz A,Morales AJ,Lomedico PT,Recker S M,Van Eerdewegh P,Recker R R,Johnson ML.Amutationinthe LDLreceptor-related protein5gene results in the autosomal dominant high-bone-mass trait.Am J Hum Genet,2002,70(1):11~19 5 Babij P,Zhao W,Small C,Kharode Y,Yaworsky PJ,Bouxsein ML,Reddy P S,Bodine P V,Robinson J A,Bhat B,Marzolf J,Moran R A,Bex F.High bone mass in mice expressing a mutantLrp5gene.J Bone Miner Res,2003,18(6):960~974 6 吕萍,龚瑶琴,李江夏,周海斌,陈丙玺.小鼠Lrp5基因基本启动子分析.山东大学学报(L櫣Ping,Gong Yao-Qin,Li Jiang-Xia,Zhou Hai-Bin,Chen Bing-Xi.Analysis of the basal promoter of mouse Lrp5gene.JShandong Univ),2004,42(4):390~394 7 吕萍,龚瑶琴,李江夏,周海斌,陈丙玺.小鼠Lrp5基因启动子的克隆及功能分析.中国生物化学与分子生物学报(L櫣Ping,Gong Yao-Qin,Li Jiang-Xia,Zhou Hai-Bin,Chen Bing-Xi.Identification of mouseLrp5gene promoter.Chin JBiochem Mol Biol),2005,21(1):72~77 8 Hey PJ,Twells R CJ,Phillips MS,Yusuke Nakagawa,Brown S D,Kawaguchi Y,Cox R,Xie G C,Dugan V,Hammond H,Metzker M L,Todd J A,Hess J F.Cloning of a novel member of the low-density lipoprotein receptor family.Gene,1998,216(1):103~111 9 Dong Y,Lathrop W,Weaver D,Qiu Q,Cini J,Bertolini D,Chen D.Molecular cloning and characterization of LR3,a novel LDL receptor family protein with mitogenic activity.Biochem Biophys Res Commun,1998,251(3):784~790 10 Chen D,Lathrop W,Dong Y.Molecular cloning of mouse Lrp7(Lrp3)cDNA and chromosomal mapping of orthologous genes in mouse and human.Genomics,1999,55(3):314~321
PDF(362 KB)

154

Accesses

0

Citation

Detail

Sections
Recommended

/