Development of Transgenic Maize using Immature Embryos of HiII Genotype as a Vaccine Candidate

Hyun A Kim,Pil Son Choi,Moon Sik Yang,Suk Yoon Kwon,Han Sang Yoo

doi:10.3844/ajbbsp.2012.21.24

Abstract

Problem statement: Plant-based vaccines posses some advantages over other types of vaccine biotechnology such as safety, low cost of m ass vaccination programs and wider use of vaccines for veterinary medicine. These study was undertaken to develop the transgenic maize as edible vaccine candidate for animals. Approach: The immature embryos of HiII genotype were inoculated with A. tumefaciens strain C58C1 containing the binary vector V622. The vector was harbored nptII gene, which confers resistance to paromomycin and ApxIIA gene was produced ApxII toxin, which was generated in various serum types of A. pleuropneumoniae as a target gene. Results: The 1,027 immature embryos were immersed for 5 min in the Agrobacterium solution and then these were co-cultured on solid co- cultivation medium at 28°C for 2 days. After the de lay period, the scutellum explants, axis removed embryos, were cultured on medium with 50 mg L -1 paromomycin for first 2 weeks and a paromomycin- resistant callus were sorted out on the selection m edium with 100 mg L -1 paromomycin for 4 ×14 days. A total of twenty callus clones were selected and six teen-putative transgenic plants were regenerated. Among them, only five plants contained the integrat ed nptII gene, which was confirmed by Southern blot analysis. Conclusion: These results demonstrated that the nptII and ApxIIA genes integrated into the maize genome and that transgenic maizes can be use as vaccine candidate.

Highlights

Transgenic plants have been used in the production the development of integration techniques into plants and delivery of edible oral subunit vaccines (Floss et genome of antigen gene had been tried, the expression al., 2007), because of some advantages, such as of its hetero-protein and of Hepatitis B surface Antigen increased safety, anticipated low cost of mass (HBsAg) have been reported in tobacco plants (Curtiss vaccination programs and wider use of vaccinations for veterinary (Shin et al, 2011)
We developed the transgenic maize plants antibiotics
After 4-5 days, scutellum explants were carefully dissected from the germinated immature embryos and were cultured on selection medium with MS salts, MS vitamins, 0.4 mg L−1 thiamine-HCL, 1 mg L−1 2,4-D, 20 mM MES, 0.115 g L−1 L-proline, 2% sucrose, 100 mg L−1 myo-inositol, 0.8% BBL agar (Gifco Ltd.), 300 mg L−1 cefotaxime sodium, 50 mg L−1 paromomycin at dark, 25°C for first two-weeks

Summary

Introduction

Transgenic plants have been used in the production the development of integration techniques into plants and delivery of edible oral subunit vaccines (Floss et genome of antigen gene had been tried, the expression al., 2007), because of some advantages, such as of its hetero-protein and of Hepatitis B surface Antigen increased safety, anticipated low cost of mass (HBsAg) have been reported in tobacco plants (Curtiss vaccination programs and wider use of vaccinations for veterinary (Shin et al, 2011). Grain seeds are appropriate candidate of transgenic maize expressing ApxIIA gene systems for the oral delivery of subunit vaccines using embryogenic calli co-cultivation transformation After 4-5 days, scutellum explants were carefully dissected from the germinated immature embryos and were cultured on selection medium (pH5.8) with MS salts, MS vitamins, 0.4 mg L−1 thiamine-HCL, 1 mg L−1 2,4-D, 20 mM MES, 0.115 g L−1 L-proline, 2% sucrose, 100 mg L−1 myo-inositol, 0.8% BBL agar (Gifco Ltd.), 300 mg L−1 cefotaxime sodium, 50 mg L−1 paromomycin at dark, 25°C for first two-weeks.

Results

Conclusion