Geri Dön

Characterization of an operon involved in mycobacterial cholesterol metabolism

Başlık çevirisi mevcut değil.

PDF İndir

  1. Tez No: 400762
  2. Yazar: NATHAN LACK
  3. Danışmanlar: PROF. EDITH SIM
  4. Tez Türü: Doktora
  5. Konular: Eczacılık ve Farmakoloji, Pharmacy and Pharmacology
  6. Anahtar Kelimeler: Belirtilmemiş.
  7. Yıl: 2009
  8. Dil: İngilizce
  9. Üniversite: University of Oxford
  10. Enstitü: Yurtdışı Enstitü
  11. Ana Bilim Dalı: Eczacılık Ana Bilim Dalı
  12. Bilim Dalı: Belirtilmemiş.
  13. Sayfa Sayısı: 215

Özet

Özet yok.

Özet (Çeviri)

In the recently identified cholesterol catabolic pathway of Mycobacterium tuberculosis, 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase (HsaD) is proposed to catalyze the hydrolysis of a C-C bond in the cholesterol metabolite 4,9-DSHA (4,5- 9,10-diseco-3-hydroxy-5,9,17-trioxoandrosta-1(10),2-diene-4-oic acid). The work in this thesis describes the cloning and expression of the hsaD gene from M. tuberculosis. Using a heterologous expression system in Psuedomonas putida and affinity chromatography, HsaD was purified to apparent homogeneity. To test the hydrolytic activity of the enzyme, the coloured substrate 2-hydroxy-6-oxo-6-phenylhexa- 2,4-dienoic acid was synthesized and a 96-well assay was developed for HsaD. With the purified enzyme and hydrolytic assay, the enzymatic and physical properties of the protein were characterized. Using the purified protein and an enzymatically inactive form of HsaD, the structure of this enzyme either alone or in complex with several different substrates, including 4,9-DSHA, was determined to 1.8-2.35Å. In addition to this work, a series of genetic knockouts of the nat/hsa operon were created in Mycobacterium smegmatis and Mycobacterium bovis BCG. Deletion of hsaD in M. smegmatis had no effect on the growth rate of the bacteria with cholesterol, while the deletion of the gene hsaC was found to cause the bacterial supernatant to turn pink when grown with cholesterol. With ?hsaD M. bovis BCG the mutant strain was found to grow much slower on minimal medium supplemented with cholesterol. Deletion of hsaD in M. bovis BCG was also found to inhibit intracellular growth in macrophages. Further to this work, using antibodies raised against proteins encoded by this operon and genetic knockouts, the nat/hsa operon was found to contain an additional promoter. Furthermore, M. bovis BCG was found to significantly alter the level and distribution of several lipid species in macrophages, including cholesterol. These effects shed light on the mechanism of mycobacterial survival inside macrophages.

Benzer Tezler

  1. Tez No

    815755

    The productıon and pegylatıon of recombinant human granulocyte colony-stımulatıng factor produced ın e. coli

    Rekombinant insan granulosit koloni stimülan faktörünün e. coli'de üretimi ve pegilasyonu

    NAZLI DİLARA ERDOĞDU

    Yüksek Lisans

    İngilizce

    İngilizce

    2023

    Biyokimyaİstanbul Teknik Üniversitesi

    Moleküler Biyoloji-Genetik ve Biyoteknoloji Ana Bilim Dalı

    PROF. DR. GİZEM DİNLER DOĞANAY

  2. Tez No

    402754

    Genome-wide prediction of prokaryotic two-component system networks using a sequence-based meta-predictor

    Başlık çevirisi yok

    ALTAN KARA

    Doktora

    İngilizce

    İngilizce

    2016

    BiyolojiAberystwyth University / Prifysgol Aberystwyth

    Yurtdışı Enstitü

    DR. NARCIS FERNANDEZ-FUENTES

    DR. DAVID WHITWORTH

  3. Tez No

    397770

    Bacillus subtilis GntR ailesine ait LutR transkripsiyon faktörünün doğrudan kontrolü altındaki genlerin CHIP ve EMSA yöntemleriyle belirlenmesi

    Determination of genes under the direct control of GntR-type transcriptional factor LutR in Bacillus subtilis PY79 by CHIP and EMSA methods

    MURAT KEMAL AVCI

    Doktora

    Türkçe

    Türkçe

    2015

    Biyoteknolojiİstanbul Teknik Üniversitesi

    Moleküler Biyoloji-Genetik ve Biyoteknoloji Ana Bilim Dalı

    PROF. DR. AYTEN KARATAŞ

  4. Tez No

    401666

    Characterization of the Pseudomonas aeruginosa mepa, a unique metalloendopeptidase whose gene is a part of the VFR regulon

    Başlık çevirisi yok

    AYŞEGÜL BALYİMEZ

    Doktora

    İngilizce

    İngilizce

    2013

    BiyolojiTexas Tech University

    Biyolojik Bilimler Ana Bilim Dalı

    DR. ABDUL HAMOOD

    DR. MICHAEL SAN FRANCISCO

  5. Tez No

    95433

    Isolation and characterization of rust resistance marker DNAs of wheat

    Buğdayda pas hastalığına karşı direnç markör DNAlarının isolasyonu ve tanımlanması

    SEVGİ SALMAN

    Yüksek Lisans

    İngilizce

    İngilizce

    2000

    BiyolojiBoğaziçi Üniversitesi

    Moleküler Biyoloji ve Genetik Ana Bilim Dalı

    DOÇ. DR. MÜGE TÜRET SAYAR

Geri Dön