Plant-based production of recombinant proteins has emerged as an efficient and cost-effective alternative to microbial fermentation and mammalian cell culture systems. Chloroplasts harbor high plasmid copy numbers and can be stably transformed, making them efficient platforms for protein production. In the present study, we used green fluorescent protein (GFP) as a reporter to compare the three major chloroplast promoters (rrn, psbA, and rbcL) involved in protein production in Nicotiana tabacum cv. “Petit Havana.” Three chloroplast transformation vectors were constructed, each regulated by a different promoter, and the transformation was performed via biolistic particle bombardment. Transformants were selected based on spectinomycin resistance and were confirmed by PCR. Among the three promoters, psbA showed the highest transformation efficiency and protein expression levels. Reverse transcription quantitative PCR showed that the mRNA levels (relative to Actin) for psbA (218.21±19.64) were nearly twice that of rbcL (126.60±8.78), and five times that of rrn (43.27±1.57). This transcriptional hierarchy was also observed at protein level. Immunoblotting showed the GFP levels (relative to psbA) were: psbA (1.00), rbcL (0.87), and rrn (0.77), whereas quantification through ELISA revealed relative GFP concentrations of: 616.2±28.7 ng/g LFW for psbA, 510.3±32.4 ng/g LFW for rbcL, and 338.9±100.2 ng/g ng/g LFW for rrn. These quantitative results demonstrate the importance of promoter selection for efficient expression of recombinant proteins in chloroplasts and show that the psbA promoter is suitable for high-efficiency chloroplast expression systems, providing a foundation for advancing plant-based molecular farming.

Wheat is a fundamental staple crop worldwide, contributing significantly to global food security due to its versatility and nutritional value. However, gluten proteins, including gliadins and glutenins, have been implicated in various health problems, such as celiac disease, non-celiac gluten sensitivity, and wheat allergies. These disorders affect a wide variety of people globally, creating demand for wheat varieties that balance high-end-use quality with reduced immunogenic potential. This review examines the molecular and genetic mechanisms that regulate gluten protein synthesis, highlighting recent advances in genomic and mutagenic approaches aimed at modifying gluten proteins to enhance the health and quality traits of wheat. Technologies such as RNAi and CRISPR/Cas9 offer promising avenues for reducing wheat immunogenicity without compromising its functional properties in food production. This study also examines the challenges and prospects of utilizing these genetic tools to develop wheat varieties that achieve the dual
objective
s of enhanced health outcomes and high product quality.