Qamar Firdaus, Khan Shazia, Ashrafi Kudsiya, Iqrar Sadia, Quadri Syed Naved, Saifi Monica, Abdin M Z
Centre for Transgenic Plant and Development, Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi,110062, India.
Biotechnol Rep (Amst). 2022 Aug 30;36:e00761. doi: 10.1016/j.btre.2022.e00761. eCollection 2022 Dec.
The therapeutic efficacy of L. is governed by artemisinin (ART), prevalently produced by extraction. Due to the modest amount of ART (0.01-1 %dw) in this plant, commercialization of ACTs is difficult. In this study, the floral-dip based transformation protocol for was developed to enhance expression of artemisinin biosynthesis genes and ART content. For dipping, the effective infiltration media components were optimized, and to obtain high transformation (26.9%) partially open bud stage capitulum of floral development was used. Hygromycin phospho-transferase () selection marker was used to validate the transformed T progenies. The copy numbers of the transgene () in T progenies were determined using a sensitive, high-throughput SYBR Green based quantitative RT-PCR. The results of the transgene were compared with those of the low copy number, internal standard (). Using optimised PCR conditions, one, two and three transgene copies in T transformants were achieved.
黄花蒿的治疗功效取决于青蒿素(ART),青蒿素主要通过提取产生。由于该植物中青蒿素的含量较低(0.01-1%干重),青蒿素联合疗法(ACTs)的商业化生产面临困难。在本研究中,开发了基于花浸染的黄花蒿转化方案,以增强青蒿素生物合成基因的表达和青蒿素含量。对于浸染,优化了有效的浸润培养基成分,并使用花发育的部分开放芽期头状花序以获得高转化率(26.9%)。潮霉素磷酸转移酶()选择标记用于验证转化的T代后代。使用基于SYBR Green的灵敏、高通量定量RT-PCR测定T代后代中转基因()的拷贝数。将转基因()的结果与低拷贝数内标()的结果进行比较。使用优化的PCR条件,在T转化体中实现了一、二和三个转基因拷贝。