New-Generation Vectors for Plant Transgenics: Methods and Applications
Transgenic development is the establishment of novel traits into the plants to enhance its quality. Foreign gene introduction into the nuclear or chloroplast genomes in plants is achieved through the DNA-carrying elements known as plasmids or vectors. Plant genetic engineering will be productive only if we develop small, easy-to-handle, and simple to use Agrobacterium binary vectors. Most of the new-generation vectors were derived from conventional vectors, such as pBIN and pCAMBIA series. Conventional vectors are larger in size making it difficult for cloning as it decreases the ability of gene integration. For the functional characterization of genes, it requires comprehensive genetic analysis, which includes overexpression, downregulation (antisense/RNAi), promoter analysis, subcellular localization studies, and gene complementation analysis. These high-throughput functional genetic approaches rely on efficient cloning strategies and new-generation vectors. Ancient cloning procedures based on the restriction and digestion are cumbersome and require large time. Modern cloning approaches were established with the newly arrived next-generation vector systems that will be helpful to reduce the cloning difficulty and to increase cloning efficiency methods. The major purpose of the expression vectors is to achieve high protein expression, which is normally driven by strong promoters. Modern, stable, and transient expression vector systems were established to enhance the expression of the foreign gene and to reduce the complexity of gene construct preparation. This chapter describes the various new-generation vectors and their potential application in the field of plant genetic engineering.
KeywordsPlasmids Vectors Cloning Promoters Gene expression Genetic engineering
This study was supported by a grant (Sanction No. PDF/2016/000750) from the Department of Science and Technology, Science and Engineering Research Board, Government of India, and (Sanction No. No.F.4-2/2006 (BSR)/BL/16-170541) from the D. S. Kothari Postdoctoral Fellowship and was also supported by UGC-SAP, DST-FIST, and DST-PURSE schemes.
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