University of Louisville
Understanding the Evolution of Pathogenic Feline Leukemia Virus C
Institution
University of Louisville
Faculty Advisor/ Mentor
Chet Tailor; Naveen Hussain
Abstract
FeLV is a member of the retrovirus family characterized by transcribing RNA into DNA using the enzyme reverse transcriptase. In a retrovirus the env domain of the 3 part retrovirus genome codes for the Surface Glycoprotein, composed of RBD, PRR, and C domain, and the Transmembrane Protein. My experiments focus on the role of the RBD, which determines the receptor specificity and the PRR which forms a hinge between RBD and the c-domain. Past research on FeLV have shown that these viruses could be more efficient vectors for gene therapy than the presently used MLV due to better infectivity they have on human hematopoietic stem cells. There are 4 subgroups of FeLV: A, B, C and T with key focus is on subgroup A, which can be transmitted from cat to cat and C is formed by mutations. FeLV-A, the most common type of the virus, uses the receptor THTR1 a thiamine transporter and FeLV-C uses FLVCR1 a heme exporter receptor. To study the evolution of pathogenic FeLV-C from FeLV-A our lab obtained a variant of FeLV-A FA27-53 with expanded host range to pigs. This project is to find whether FA27-54 uses an alternate receptor by determining the receptor(s) to infect pig cells. Using PCR the envelopes of FeLV-A 61E, FA27-53 and FeLV-A CL-29 were isolated and inserted successfully into a pFBNeo vector. These plasmids were then used to transfect CCC feline and StIowa pig cell lines and treated with G418.
Understanding the Evolution of Pathogenic Feline Leukemia Virus C
FeLV is a member of the retrovirus family characterized by transcribing RNA into DNA using the enzyme reverse transcriptase. In a retrovirus the env domain of the 3 part retrovirus genome codes for the Surface Glycoprotein, composed of RBD, PRR, and C domain, and the Transmembrane Protein. My experiments focus on the role of the RBD, which determines the receptor specificity and the PRR which forms a hinge between RBD and the c-domain. Past research on FeLV have shown that these viruses could be more efficient vectors for gene therapy than the presently used MLV due to better infectivity they have on human hematopoietic stem cells. There are 4 subgroups of FeLV: A, B, C and T with key focus is on subgroup A, which can be transmitted from cat to cat and C is formed by mutations. FeLV-A, the most common type of the virus, uses the receptor THTR1 a thiamine transporter and FeLV-C uses FLVCR1 a heme exporter receptor. To study the evolution of pathogenic FeLV-C from FeLV-A our lab obtained a variant of FeLV-A FA27-53 with expanded host range to pigs. This project is to find whether FA27-54 uses an alternate receptor by determining the receptor(s) to infect pig cells. Using PCR the envelopes of FeLV-A 61E, FA27-53 and FeLV-A CL-29 were isolated and inserted successfully into a pFBNeo vector. These plasmids were then used to transfect CCC feline and StIowa pig cell lines and treated with G418.