J 2002

BAC library of T. pallidum DNA in E. coli.

ŠMAJS, David, M. MCKEVITT, L. WANG, J.K. HOWELL, S.J. NORRIS et. al.

Basic information

Original name

BAC library of T. pallidum DNA in E. coli.

Authors

ŠMAJS, David, M. MCKEVITT, L. WANG, J.K. HOWELL, S.J. NORRIS, T. PALZKILL and G.M. WEINSTOCK

Edition

Genome Res. 2002

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

Organization unit

Faculty of Medicine

UT WoS

000174171300019
Změněno: 25/6/2009 15:47, prof. MUDr. David Šmajs, Ph.D.

Abstract

V originále

Treponema pallidum subspecies pallidum (Nichols) chromosomal DNA was used to construct a large insert bacterial artificial chromosome (BAC) library in Escherichia coli DH10B using the pBeloBAC11 cloning vector; 678 individual insert termini of 339 BAC clones (13.9 x coverage) were sequenced and the cloned chromosomal region in each clone was determined by comparison to the genomic sequence. A single 15.6-kb region of the T. pallidum chromosome was missing in the BAC library, between bp 248727 and 264323. In addition to the 12 open reading frames (ORFs) coded by this region, one additional ORF (TP0596) was not cloned as an intact gene. Altogether, 13 predicted T. pallidum ORFs (1.25% of the total) were incomplete or missing in the library. Three of 338 clones mapped by restriction enzyme digestion had detectable deletions and one clone had a detectable insertion within the insert. Of mapped clones, 19 were selected to represent the minimal set of E. coli BAC clones covering 1026 of the total 1040 (98.7%) predicted T. pallidum ORFs. Using this minimal set of clones, at least 12 T. pallidum proteins were shown to react with pooled sera from rabbits immunized with T. pallidum, indicating that at least some T. pallidum genes are transcribed and expressed in E. coli.