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In Vitro Generation Of: Immortalized CD34+

The document describes a study that aimed to generate immortalized CD34+ haematopoietic stem cells by transducing them with the HPV16-E6/E7 gene using a lentiviral vector system. The HPV16-E6/E7 gene was inserted into the pLVX-TetOne-Puro plasmid and transfected into CD34+ cells. Transduced cells were selected for puromycin resistance and analyzed using qRT-PCR and western blot to confirm HPV16-E6/E7 expression. The immortalized CD34+ cells could be differentiated into various blood cell types.

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37 views3 pages

In Vitro Generation Of: Immortalized CD34+

The document describes a study that aimed to generate immortalized CD34+ haematopoietic stem cells by transducing them with the HPV16-E6/E7 gene using a lentiviral vector system. The HPV16-E6/E7 gene was inserted into the pLVX-TetOne-Puro plasmid and transfected into CD34+ cells. Transduced cells were selected for puromycin resistance and analyzed using qRT-PCR and western blot to confirm HPV16-E6/E7 expression. The immortalized CD34+ cells could be differentiated into various blood cell types.

Uploaded by

Mind Niramind
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as ZIP, PDF, TXT or read online on Scribd
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In vitro generation of

Immortalized CD34+
Haematopoietic Stem Cells
with HPV16-E6/E7 gene

Presented by
Presented by Ms.
Ms. Niramol
Niramol Prombandit
Prombandit 5936312
5936312
SCBT/M
SCBT/M
Biotechnology, Science,
Biotechnology, Science, Mahidol
Mahidol University
University
Advisor:Dr. Suparerk
Advisor:Dr. Suparerk Borwornpinyo
Borwornpinyo
Co-advisor: Dr.
Co-advisor: Dr. Lalana
Lalana Nuntakarn
Nuntakarn
2
1 pLVX-TetOneTM-Puro 3
HPV16-E6/E7 gene Transient
transfection

•
qRT-PCR
•
WESTERN BLOT

MATERIALS & METHO


Lentivirus

CD34+ HSC

6
Cell Differentiation 5
Immortal 4
Lentiviral
Check Ligation
Check Ligation

Plasmid
Plasmid
Extraction
Extraction

Digest

BshTI
XhoI

TM
pLVX-TetOneTM
pLVX-TetOne -Puro++HPV16-E6/E7
-Puro HPV16-E6/E7
LB Agar

Plasmid Result
Result
Plasmid Report
Extraction
Extraction Report

DNA
TM
pLVX-TetOneTM -Puro
pLVX-TetOne -Puro
++HPV16-E6/E7
HPV16-E6/E7 DNA
Alignmen
Alignmen
tt

Colonies
Colonies
PCR
PCR
HPV16-E6/E7
HPV16-E6/E7

Sequencin
Sequencin
gg Analysis
Analysis

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