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Ahmed, Golam Rizvee.. Investigation of halotag fusion protein function in E. coli and plasmodium. Master's Degree(Biochemistry). Mahidol University. Mahidol University Library and Knowledge Center. : Mahidol University, 2014.
Investigation of halotag fusion protein function in E. coli and plasmodium
Name: Ahmed, Golam Rizvee.
LCSH:
Protein engineering.
Abstract:
HaloTag (HT) is a modified bacterial haloalkane dehalogenase enzyme that has been developed as a multi-purpose tool for studying protein functions, including protein purification, intracellular protein localization, time-dependent labeling, and control of intracellular protein levels. Ligand-mediated control of intracellular protein is an attractive tool for drug target validation. The function of HT was initially tested in E. coli. EGFP-HT fusion protein was functional when expressed in E. coli, although the expression was markedly lower than expected. Another reporter protein, dihydrofolate reductase thymidylate synthase (DHFR-TS) was used as a fusion partner to HT and its function was tested in E. coli. It was observed that the fusion of HT to DHFR-TS protein lowered the expression and intracellular activity of DHFR-TS compared with a 6x His tagged DHFR-TS. The DHFRTS-HT fusions had correspondingly lower DHFR activity to complement a DHFR-TS E. coli mutant. The expression of the EGFP-HT fusion protein was also tested in the blood stages of the Plasmodium berghei rodent malaria parasite. Correct integrants were validated by PCR. EGFP-HT mRNA expression was detected by RT-PCR in transgenic EGFP-HT parasite lines. However, no EGFP-HT fusion protein was detected by Western blot. In conclusion, HT can be expressed as a fusion protein, but the low levels of protein activity suggest that HT interferes with fusion protein expression and/or stability in E. coli and Plasmodium sp."
Mahidol University. Mahidol University Library and Knowledge Center
Address:
NAKHONPATHOM
Email:
liwww@mahidol.ac.th
Name: Thanat Chookajorn
Role:
Thesis Advisors
Name: Ornchuma Itsathiphaisarn
Role:
Thesis Advisors
Name: Shaw, Philip James
Role:
Thesis Advisors
Created:
2014
Modified:
2020-04-14
Issued:
2019-11-07
วิทยานิพนธ์/Thesis
application/pdf
CallNumber:
TH A286i 2014
eng
DegreeName: Master of Science
Level: Master's Degree
Descipline: Biochemistry
Grantor: Mahidol University
©copyrights Mahidol University
RightsAccess:
ใช้เวลา
0.377679 วินาที
Ahmed, Golam Rizvee.
| Title | Contributor | Type |
|---|---|---|
| Investigation of halotag fusion protein function in E. coli and plasmodium
มหาวิทยาลัยมหิดล Ahmed, Golam Rizvee. | Thanat Chookajorn Ornchuma Itsathiphaisarn Shaw, Philip James | วิทยานิพนธ์/Thesis |
Thanat Chookajorn
| Title | Creator | Type and Date Create |
|---|---|---|
| Investigation of halotag fusion protein function in E. coli and plasmodium
มหาวิทยาลัยมหิดล Thanat Chookajorn;Ornchuma Itsathiphaisarn;Shaw, Philip James | Ahmed, Golam Rizvee. | วิทยานิพนธ์/Thesis |
Ornchuma Itsathiphaisarn
| Title | Creator | Type and Date Create |
|---|---|---|
| Investigation of halotag fusion protein function in E. coli and plasmodium
มหาวิทยาลัยมหิดล Thanat Chookajorn;Ornchuma Itsathiphaisarn;Shaw, Philip James | Ahmed, Golam Rizvee. | วิทยานิพนธ์/Thesis |
Shaw, Philip James
| Title | Creator | Type and Date Create |
|---|---|---|
| Investigation of halotag fusion protein function in E. coli and plasmodium
มหาวิทยาลัยมหิดล Thanat Chookajorn;Ornchuma Itsathiphaisarn;Shaw, Philip James | Ahmed, Golam Rizvee. | วิทยานิพนธ์/Thesis |