NR APMJ
AU Sun,X.; Dong,X.; Zhou,W.
TI [The sequence of the human PrP gene Exon I and its upstream segment has promoter-like activity]
QU Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese Journal of Experimental and Clinical Virology 2000 Dec; 14(4): 305-8
PT journal article
AB OBJECTIVE: The human PrP gene locates at the short-arm of the 20th chromosome. This article is was to map the promoter that transcribes the human PrP gene. METHODS: The sequence of the human PrP gene Exon I and its upstream segment were amplified using PCR, and inserted into a CAT reporter plasmid pBL-CAT6 after sequence analysis. The values of the relative CAT expression under the control of this fragment were evaluated after transiently transfected into HeLa, COS7 and Sh-sy5y cell lines. The amounts of transcription activator SP1 in these three cell lines were calculated with band-shift assays. RESULTS: Analysis of the human PrP gene Exon I and its upstream segment showed a GC rich sequence, with several potential SP1 binding sites, but without any TATA-box. Under the control of this fragment, the CAT expressions were 2-3 folds increased in transient transfection. Quantity band-shift assays revealed that SP1 was enriched in HeLa cells, but undetectable in COS7 and Sh-sy5y cells. CONCLUSIONS: The sequence of the human PrP gene Exon I and its upstream segment functions as a promoter-like sequence, probably being as a weak TATA-less promoter. Cells derived from different tissues contain different amount of SP1. The activity of this promoter-like sequence seems to be independent of SP1 presence.
MH Cell Line; Exons/*genetics; Human; Plasmids; Polymerase Chain Reaction; Prions/*genetics; Promoter Regions (Genetics)/*genetics; Support, Non-U.S. Gov't; Transcription Factor, Sp1/analysis; Transcription Initiation Site; Transcription, Genetic; Transfection
AD Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052, China.
SP englisch
PO China