Expression of Protein Antigen Hiv -1 P24 in Transgenic Tobacco Plants
VIETNAM NATIONAL UNIVERSITY – HOCHIMINH CITY
INTERNATIONAL UNIVERSITY
School of Biotechnology
EXPRESSION OF PROTEIN ANTIGEN HIV -1 P24 IN TRANSGENIC TOBACCO PLANTS
A thesis submitted to
The School of Biotechnology, International University
In partial fulfillment of the requirements for the degree of
B.S. in Biotechnology
Student name: Nguyen Ngoc Tu Anh – BT050036 Supervisor: 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 traditional production systems. Transgene expression from the plant’s plastid genome represents a promising strategy in molecular farming because of the plastid’s potential to accumulate foreign proteins to high levels and the increased biosafety provided by the maternal mode of organelle inheritance. In this thesis, we confirm the high-level expression of HIV-1 p24 antigen in transgenic tobacco plants through plastid transformation. PCR analysis confirmed the presence of the HIV-1 p24 sequence within the chloroplast genome of transgenic lines. SDS-PAGE gel electrophoresis of protein extracts from transgenic plants identified plant-expressed HIV-1 p24 protein. Quantification of the recombinant protein HIV-1 p24 using Aligent 2100 Bioanalyzer estimated yields of about 13 mg per g of soluble leaf protein. Our results indicate that plant-based transgenic expression represents the great potential of transgenic plastids to develop HIV vaccine components at low cost and high yield and for use in HIV diagnostic procedures.
ACKNOWLEDGMENT
First and foremost, I would like to thank to Dr. Nguyen Thi Thanh for the valuable guidance and advice to complete my thesis for Bachelor of Science degree. I also would like to thank to Dr. Phan Dinh Phap for his help and introduction to the Institute of Tropical Biology.
INTERNATIONAL UNIVERSITY
School of Biotechnology
EXPRESSION OF PROTEIN ANTIGEN HIV -1 P24 IN TRANSGENIC TOBACCO PLANTS
A thesis submitted to
The School of Biotechnology, International University
In partial fulfillment of the requirements for the degree of
B.S. in Biotechnology
Student name: Nguyen Ngoc Tu Anh – BT050036 Supervisor: 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 traditional production systems. Transgene expression from the plant’s plastid genome represents a promising strategy in molecular farming because of the plastid’s potential to accumulate foreign proteins to high levels and the increased biosafety provided by the maternal mode of organelle inheritance. In this thesis, we confirm the high-level expression of HIV-1 p24 antigen in transgenic tobacco plants through plastid transformation. PCR analysis confirmed the presence of the HIV-1 p24 sequence within the chloroplast genome of transgenic lines. SDS-PAGE gel electrophoresis of protein extracts from transgenic plants identified plant-expressed HIV-1 p24 protein. Quantification of the recombinant protein HIV-1 p24 using Aligent 2100 Bioanalyzer estimated yields of about 13 mg per g of soluble leaf protein. Our results indicate that plant-based transgenic expression represents the great potential of transgenic plastids to develop HIV vaccine components at low cost and high yield and for use in HIV diagnostic procedures.
ACKNOWLEDGMENT
First and foremost, I would like to thank to Dr. Nguyen Thi Thanh for the valuable guidance and advice to complete my thesis for Bachelor of Science degree. I also would like to thank to Dr. Phan Dinh Phap for his help and introduction to the Institute of Tropical Biology.
References: | Hepatitis B | (Huang et al,2005) | Cholera | (Jani et al, 2004; Mishra et al, 2006) | Rotavirus | (Birch-Machin et al, 2004; Perez Filgueira et al, 2004) | Human papillomavirus | (Wen et al, 2006), (Massa et al, 2007; Fernandez-San Millan et al, 2008; Lenzi et al, 2008) | Foot and mouth disease (FMDV) | (Wigdorovitz et al, 1999; Wu et al Diabetes | (Avesani et al, 2003) | SARS | (Li et al, 2006) | Tuberculosis | (Zelada et al, 2006; Dorokhov et al, 2007) | Newcastle disease virus | (Zhao and Hammond, 2005) | Smallpox | (Golovkin et al, 2007) | Poliovirus | (Fujiyama et al, 2006) | Lyme disease | (Navarre et al, 2006) | FMDV and hepatitis B | (Huang et al, 2005) | HIV and rabies | (Yusibov et al, 1997) | 2.2 HUMAN IMMUNODEFICIENCY VIRUS TYPE 1(HIV-1) From (Lindh, 2009) 2.2.2 Replication cycle of HIV-1