Link to the University of Pittsburgh Homepage
Link to the University Library System Homepage Link to the Contact Us Form

Expression, purification and characterization of bacteriophage lambda tail tip proteins

Dai, Xiaoxian (2009) Expression, purification and characterization of bacteriophage lambda tail tip proteins. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

[img]
Preview
PDF
Primary Text

Download (6MB) | Preview

Abstract

Bacteriophage tails, despite differences in their morphology, all play a key role in host recognition and DNA injection. It is widely believed that the tail, especially the baseplate / tail tip, has to undergo conformational change and protein rearrangement during infection. This change has been observed in both the long, contractile tail of T4 and the short tail of T7. In contrast, little is known about this aspect of the long, non-contractile tail of bacteriophage Ć. Four proteins are involved in the Ć tail tip assembly. I present evidence that gpJ, gpI, gpL and gpK are components of the tail tip complex. My results also suggest that there may be about three copies of gpI and three copies of gpL involved in the Ć tail assembly. In addition, I have successfully purified gpL, which contains eight cysteines. My results show that when the conserved cysteine at position 173, 182 or 205 is mutated to serine, the mutant protein is defective in tail assembly. However, the C212S mutant accumulates a small amount of tail. Further analysis of this mutant indicates that C212 may have roles in both tail assembly and DNA injection.gpK is required for Ć tail assembly, but is not detected in the mature virion. Two different amber mutations were introduced into gene K. Neither of these mutants is able to complement in vivo. However, the short amber fragment is unable to assemble Ć tail whereas phage-like particles with little infectivity accumulate in the long amber fragment lysate. The results indicate that the function of gene K can be bypassed to some extent in the Kam768 mutant, but not in the Kam6 mutant.


Share

Citation/Export:
Social Networking:
Share |

Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Dai, Xiaoxianxiaoxiandai@gmail.com
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairHendrix, Roger Wrhx@pitt.eduRHX
Committee MemberHatfull, Graham Fgfh@pitt.eduGFH
Committee MemberPipas, James Mpipas@pitt.eduPIPAS
Committee MemberBrodsky, Jeffrey Ljbrodsky@pitt.eduJBRODSKY
Committee MemberBerget, Peter Bberget@andrew.cmu.edu
Date: 30 September 2009
Date Type: Completion
Defense Date: 17 June 2009
Approval Date: 30 September 2009
Submission Date: 5 August 2009
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Institution: University of Pittsburgh
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: bacteriophage lambda; tail proteins
Other ID: http://etd.library.pitt.edu/ETD/available/etd-08052009-211724/, etd-08052009-211724
Date Deposited: 10 Nov 2011 19:57
Last Modified: 15 Nov 2016 13:48
URI: http://d-scholarship.pitt.edu/id/eprint/8919

Metrics

Monthly Views for the past 3 years

Plum Analytics


Actions (login required)

View Item View Item