東華大學圖書館 |

語系: 繁體中文

說明(常見問題)

回圖書館首頁

手機版館藏查詢

登入

回首頁

切換: 標籤 | MARC模式 | ISBD

Transcription study of the Escherich...

Pratt, Timothy S.

FindBook

Google Book

Amazon

博客來

Transcription study of the Escherichia coli fis promoter: Characterization of factors affecting transcription in vitro and examination of alternative transcription initiation sites in vitro and in vivo.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Transcription study of the Escherichia coli fis promoter: Characterization of factors affecting transcription in vitro and examination of alternative transcription initiation sites in vitro and in vivo./
作者:	Pratt, Timothy S.
面頁冊數:	154 p.
附註:	Source: Dissertation Abstracts International, Volume: 66-01, Section: B, page: 0117.
Contained By:	Dissertation Abstracts International66-01B.
標題:	Biology, Molecular. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3159692
ISBN:	9780496928040

Transcription study of the Escherichia coli fis promoter: Characterization of factors affecting transcription in vitro and examination of alternative transcription initiation sites in vitro and in vivo.
Pratt, Timothy S.

Transcription study of the Escherichia coli fis promoter: Characterization of factors affecting transcription in vitro and examination of alternative transcription initiation sites in vitro and in vivo. - 154 p.

Source: Dissertation Abstracts International, Volume: 66-01, Section: B, page: 0117.

Thesis (Ph.D.)--State University of New York at Albany, 2005.

This work focused on a characterization of the conditions required for in vitro transcription from the Escherichia coli fis promoter (fisP). A reproducible and reliable procedure for studying transcription from fisP in vitro was developed. fisP transcription in vitro is shown to be very sensitive to the concentrations of KCI and K-glutamate, suggesting a relatively weak RNA polymerase (RNAP)-fisP association. These initiation complexes formed in linear templates exhibit a short half-life of about 9 minutes. Three factors affect the stability of the RNAP-fisP interactions: (1) The concentration and nature of the salt utilized; KCI was even less tolerated than K-glutamate, (2) DNA supercoiling, and (3) the availability of high concentrations of the initiation nucleotide CTP. DNA supercoiling increased both the salt tolerance and transcription from fisP, and also stabilized the fisP-RNA polymerise interaction by roughly 12-fold based on half-life determinations. CTP was uniquely capable of increasing transcription from fisP by over 12-fold. It also increased the half-lives of initiation complexes on both linear and supercoiled templates. Thus, we find that CTP and DNA supercoiling work independently to stabilize initiation complexes at fisP and their combined effects may overcome that destabilizing effects of salts in vitro and in vivo.

ISBN: 9780496928040Subjects--Topical Terms:

1017719
Biology, Molecular.

Transcription study of the Escherichia coli fis promoter: Characterization of factors affecting transcription in vitro and examination of alternative transcription initiation sites in vitro and in vivo.
LDR:03407nmm 2200289 4500 001 1823850
005 20061128084353.5
008 130610s2005 eng d
020 $a 9780496928040
035 $a (UnM)AAI3159692
035 $a AAI3159692
040 $a UnM $c UnM
100 1 $a Pratt, Timothy S. $3 1912945
245 1 0 $a Transcription study of the Escherichia coli fis promoter: Characterization of factors affecting transcription in vitro and examination of alternative transcription initiation sites in vitro and in vivo.
300 $a 154 p.
500 $a Source: Dissertation Abstracts International, Volume: 66-01, Section: B, page: 0117.
500 $a Adviser: Robert Osuna.
502 $a Thesis (Ph.D.)--State University of New York at Albany, 2005.
520 $a This work focused on a characterization of the conditions required for in vitro transcription from the Escherichia coli fis promoter (fisP). A reproducible and reliable procedure for studying transcription from fisP in vitro was developed. fisP transcription in vitro is shown to be very sensitive to the concentrations of KCI and K-glutamate, suggesting a relatively weak RNA polymerase (RNAP)-fisP association. These initiation complexes formed in linear templates exhibit a short half-life of about 9 minutes. Three factors affect the stability of the RNAP-fisP interactions: (1) The concentration and nature of the salt utilized; KCI was even less tolerated than K-glutamate, (2) DNA supercoiling, and (3) the availability of high concentrations of the initiation nucleotide CTP. DNA supercoiling increased both the salt tolerance and transcription from fisP, and also stabilized the fisP-RNA polymerise interaction by roughly 12-fold based on half-life determinations. CTP was uniquely capable of increasing transcription from fisP by over 12-fold. It also increased the half-lives of initiation complexes on both linear and supercoiled templates. Thus, we find that CTP and DNA supercoiling work independently to stabilize initiation complexes at fisP and their combined effects may overcome that destabilizing effects of salts in vitro and in vivo.
520 $a The second part of the work involved a rigorous analysis of alternative promoters upstream of fisP that have been reported to significantly contribute to the transcription of the fis operon. Our evidence, however, argues strongly for a single promoter, fisP, that is responsible fully responsible for the expression and regulation of the fis operon. This is supported by in vivo transcription experiments and by three independent assays to examine fis promoter activity in vivo: primer extension assays, S-I nuclease assays, and beta-galactosidase assays. Finally, we provide convincing evidence to demonstrate that the previously reported the alternative transcription initiation sites obtained by primer extension assays are artifacts resulting from oligonucleotide mispriming events (mostly at ribosomal RNAs) that efficiently extend when very high quantities of reverse transcriptase enzyme and relatively low temperature annealing conditions are used, as can be incorrectly recommended by the vendor.
590 $a School code: 0668.
650 4 $a Biology, Molecular. $3 1017719
650 4 $a Biology, Microbiology. $3 1017734
690 $a 0307
690 $a 0410
710 2 0 $a State University of New York at Albany. $3 769258
773 0 $t Dissertation Abstracts International $g 66-01B.
790 1 0 $a Osuna, Robert, $e advisor
790 $a 0668
791 $a Ph.D.
792 $a 2005
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3159692