DNA insulator minimizing interactions between adjacent transgenes, transcriptomes contrasting callus induction of somatic embryogenesis in maize B73
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[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT AUTHOR'S REQUEST.] In plants, transgene expression and inheritance patterns are either in some cases unpredictable or in most cases vary between different transgenic events. Transgene expression can be influenced by the sequences within transgene constructs and the chromosomal position effect. To minimize the influence between adjacent genes, one possibly effective method would be the use of a genetic insulator between adjacent genes. The palindromic insulator is one of the few plant insulators that has been studied. Here we designed and tested several synthetic palindromic insulators derived from MARs (matrix attachment region sequence) with repeated sequences that can form hairpin structures. The results show that these synthetic insulators have good utility to minimize the interactions between the adjacent genes. Maize is widely cultivated throughout the world as an important crop. Genetic transformation is an important tool in maize research and genetic improvement. Nevertheless, there's still no report of successful B73 transformation or regeneration to date. The bottleneck has been the lack of a somatic embryogenesis system in B73. Here we have applied RNA-Seq to reveal potential candidate genes involved in somatic embryogenesis in maize inbred B73 and to provide transcriptome landscape of cells during maize B73 somatic embryogenesis process. Genes that were expressed differentially between embryogenic and nonembryogenic calli were screened and transcript levels of some top candidate genes were validated through real-time PCR. Gene clusters, especially transcription factors and kinase gene clusters, were identified as potential key genes that could promote maize somatic embryogenesis.
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