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Part Collection BBa_K3812008 is used to overexpress the DHAR gene in Pichia pastoris. DHAR is widely present in the chloroplast, mitochondria and cytoplasm of plants. It is an important enzyme in the AsA-GSH cycle, which catalyzes the reduction of dehydroascorbic acid (DHA) by GSH and reduces DHA to AsA. It plays an important role in maintaining the reducing ability of ascorbic acid in plants. Transgenic plants overexpressing DHAR have reduced oxidative metabolism-related enzyme activities, increased chlorophyll content, and increased photosynthetic efficiency in an ozone environment, while transgenic plants that inhibit DHAR expression have the opposite result.
In order to better optimize BBa_K3812008 so that DHAR can be better expressed, with the help of Ailurus^TM, we connect BBa_K3812001 to 2500 expression vectors and introduce them into DH5αfor expression. Finally, we screened out the best expression system for DHAR-BBa_K3812016. BBa_K3812016 contains Promoter-BBa_J56015, RBS-BBa_K3812011, 6xHis-tag-BBa_K3812012, DHAR coding Sequence-BBa_K3812001, Terminator-BBa_K3812013, Plasmid backbone-BBa_K3812014.
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Fig.1 The expression level of DHAR protein when BBa_K3812001 is connected to 2500 expression vectors