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Phrase of Protein Antigen Hiv -1 P24 in Transgenic Tobacco Vegetation

VIETNAM NATIONAL UNIVERSITY OR COLLEGE – HOCHIMINH CITY

WORLDWIDE UNIVERSITY

University of Biotechnology

EXPRESSION OF PROTEIN ANTIGEN HIV -1 P24 IN TRANSGENIC CIGARETTE PLANTS

A thesis submitted to

The School of Biotechnology, International College or university

In partial fulfillment in the requirements for the degree of W. S. in Biotechnology

Student brand: Nguyen Ngoc Tu Anh – BT050036

Boss: Dr . Nguyen Thi Thanh

Dr . Phan Dinh Phap

June/2010

ABSTRACT

The production of antigens for vaccines in plants has the potential as a safe and cost-effective alternative to classic production systems. Transgene expression from the plant's plastid genome represents a good strategy in molecular farming because of the plastid's potential to build up foreign healthy proteins to high levels as well as the increased biosafety provided by the maternal method of organelle inheritance. With this thesis, we all confirm the high-level expression of HIV-1 p24 antigen in transgenic smoking cigarettes plants through plastid alteration. PCR evaluation confirmed the existence of the HIV-1 p24 series within the chloroplast genome of transgenic lines. SDS-PAGE solution electrophoresis of protein components from transgenic plants determined plant-expressed HIV-1 p24 healthy proteins. Quantification in the recombinant protein HIV-1 p24 using Aligent 2100 Bioanalyzer estimated brings of about 13 mg every g of soluble leaf protein. Each of our results suggest that use of plant transgenic expression represents the great potential of transgenic plastids to develop HIV vaccine components at affordable and high yield and then for use in HIV diagnostic methods.

ACKNOWLEDGMENT

To start with, I would like to thank to Dr . Nguyen Thi Thanh for the valuable assistance and guidance to full my thesis for Bachelors of Research degree. I actually also would like to thank to Dr . Phan Dinh Phap for his help and introduction to the Institute of Tropical Biology. Besides, I would really like to thank the authority of the Institute of Warm Biology for providing me personally with a good environment and establishments to finish this thesis. I as well would like to give thanks to to Doctor Nguyen Huu Ho, Mister. Le Color Duc, Mister. Loc, Mister. Tri, Mister. Duong and particularly Mr. Huan who allowed me to a lot in doing experiments and gave me advantageous conditions to complete my own thesis. In addition , I would like to acknowledge the School of Biotechnology International College or university for giving me facilitates during the time I studied in this article. Finally, an honorable talk about goes to the families and friends for their understandings and supports about us in completing this kind of thesis. With out helps in the particular that mentioned above, I would personally face many difficulties although doing this thesis.

TABLE OF CONTENTS

ABSTRACTi

ACKNOWLEDGMENTii

LIST OF TABLESvi

SET OF FIGURESvii

ABBREVIATIONSviii

CHAPTER 1 ) INTRODUCTION2

SECTION 2 . MATERIALS REVIEW5

2 . 1TOBACCO PLANTS5

2 . 1 ) 1Geographical syndication of cigarettes plants5

2 . 1 . 2Botanical description of tobacco plants6

2 . 1 ) 2 . 1Stems6

2 . 1 ) 2 . 2Leaves6

2 . 1 . 2 . 3Flowers7

2 . 1 . 2 . 4Roots7

2 . 1 ) 2 . 5Seeds7

2 . 1 . 3Roles of tobacco plant life in the life8

2 . 2HUMAN IMMUNODEFICIENCY DISEASE TYPE 1(HIV-1)10

2 . 2 . 1Structure and genome of HIV-110

2 . 2 . 2Replication cycle of HIV-112

installment payments on your 2 . 3The potential of the p24 capsid protein as being a vaccine antigen13

2 . 2 . 4Facts of HIV in Viet Nam15

2 . 3PLASTID TRANSFORMATION16

2 . 3. 1Plastid and their genome structure16

2 . 3. 2Advantages and disadvantages of plastid modification over indivisible transformation16

2 . 4MOLECULAR FARMING - PLANT LIFE AS APPEARANCE SYSTEM18

2 . 5SDS-PAGE AND PROTEIN ASSAY20

2 . your five. 1Protein assay theory20

installment payments on your 5. 2Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE)20

installment payments on your 6BIOANALYZER AND PROTEIN 50 ASSAY22

2 . 6. 1Agilent 2100 bioanalyzer22

2 . 6th. 2Protein 50 LabChip kit23

CHAPTER a few. MATERIAL AND METHOD26...

References:

Hepatitis B | (Huang et ing, 2005)

Cholera | (Jani ou al, 2004; Mishra ain al, 2006)

Rotavirus | (Birch-Machin et 's, 2004; Perez Filgueira ou al, 2004)

Human papillomavirus | (Wen ain al, 2006), (Massa et al, 3 years ago; Fernandez-San Millan et 's, 2008; Lenzi et al, 2008)

Foot and mouth area disease (FMDV) | (Wigdorovitz ou al, 99; Wu ou al

Diabetes | (Avesani et al, 2003)

SARS | (Li et approach, 2006)

Tuberculosis | (Zelada et al, 06\; Dorokhov ain al, 2007)

Newcastle disease virus | (Zhao and Hammond, 2005)

Smallpox | (Golovkin et al, 2007)

Poliovirus | (Fujiyama et 's, 2006)

Lyme disease | (Navarre et ing, 2006)

FMDV and hepatitis B | (Huang et 's, 2005)

HIV and rabies | (Yusibov et al, 1997)

2 . two HUMAN IMMUNODEFICIENCY VIRUS TYPE 1(HIV-1)

Via (Lindh, 2009)

2 . 2 . 2 Duplication cycle of HIV-1

28.08.2019

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