Abstract
It has already been demonstrated that a betasatellite associated with cotton leaf curl Multan virus (CLCuMB) can be used as a plant and animal gene delivery vector to plants. To examine the ability of CLCuMB as a tool to transfer coat protein genes of HIV-1 to plants, two recombinant CLCuMB constructs in which the CLCuMB βC1 ORF was replaced with two HIV-1 genes fractions including a 696 bp DNA fragment related to the HIV-1 p24 gene and a 1501 bp DNA fragment related to the HIV-1 gag gene were constructed. Gag is the HIV-1 coat protein gene and p24 is a component of the particle capsid. Gag and p24 are used for vaccine production. Recombinant constructs were inoculated to Nicotiana glutinosa and N. benthamiana plants in the presence of an Iranian isolate of Tomato yellow leaf curl virus (TYLCV-[Ab]) as a helper virus. PCR analysis of inoculated plants indicated that p24 gene was successfully replicated in inoculated plants, but the gag gene was not. Real-time PCR and ELISA analysis of N. glutinosa and N. benthamiana plants containing the replicative forms of recombinant construct of CLCuMB/p24 indicated that p24 was expressed in these plants. This CLCuMB-based expression system offers the possibility of mass production of recombinant HIV-1 p24 protein in plants.
Highlights
Biotechnology methods using plants as an expression system offer the possibility of producing recombinant proteins for medical or veterinary applications [1, 2]
To assay the possibility of using CLCuMB as an Human Immunodeficiency Virus type 1 (HIV-1) coat protein (CP) gene expression vector, two recombinant constructs were produced by the insertion of p24 and gag genes of HIV1 into pBin-1.2βΔC1 to generate pBinβΔC1/p24 and pBinβΔC1/gag, respectively
Plant viral vectors are widely used for transient expression in plants [7, 47, 48]
Summary
Biotechnology methods using plants as an expression system offer the possibility of producing recombinant proteins for medical or veterinary applications [1, 2]. Plants have many advantages over other expression systems for producing recombinant proteins. Yeast and mammalian and insect cell cultures, plants have very low production costs, very high scale-up potential, and require low initial investments [3]. The use of plants and cell cultures to produce recombinant proteins, termed “molecular farming” [4], offers a great potential for producing high-value, cheaper, faster and widely available pharmaceuticals. In the case of viral vaccines, in particular, it has been proven that fully-assembled complex virus like.
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