Plant Biotechnology, Department of Genetics and Plant Breeding, Banaras Hindu University, Mirzapur, India.
Unit of Teaching Veterinary Clinical Complex, Faculty of Veterinary and Animal Sciences, Banaras Hindu University, Mirzapur, India.
Prep Biochem Biotechnol. 2024 Aug;54(7):882-895. doi: 10.1080/10826068.2023.2297692. Epub 2024 Jan 3.
In growing plant population, effect of stress is a perturb issue affecting its physiological, biochemical, yield loss and developmental growth. Protein-L-isoaspartate-O-methyltransferase (PIMT) is a broadly distributed protein repair enzyme which actuate under stressful environment or aging. Stress can mediate damage converting protein bound aspartate (Asp) residues to isoaspartate (iso-Asp). This spontaneous and deleterious conversion occurs at an elevated state of stress and aging. Iso-Asp formation is associated with protein inactivation and compromised cellular survival. PIMT can convert iso-Asp back to Asp, thus repairing and contributing to cellular survival. The present work describes the isolation, cloning, sequencing and expression of PIMT genes of () and () Using gene specific primers, both the were amplified from their respective cDNAs and subsequently cloned in prokaryotic expression vector pProEXHTa. BL21(DE3) strain of cells were used as expression host. The expression kinetics of both the PIMTs were studied with various concentrations of IPTG and at different time points. Finally, the PIMT supplemented BL21(DE3) cells were evaluated against different stresses in comparison to their counterparts with the empty vector control.
在植物种群的生长过程中，胁迫的影响是一个干扰问题，影响其生理、生化、产量损失和发育生长。蛋白-L-异天冬氨酸-O-甲基转移酶（PIMT）是一种广泛分布的蛋白修复酶，在胁迫环境或衰老下发挥作用。胁迫可以介导损伤，将蛋白结合的天冬氨酸（Asp）残基转化为异天冬氨酸（iso-Asp）。这种自发的、有害的转化发生在应激和衰老的高状态下。iso-Asp 的形成与蛋白失活和细胞存活受损有关。PIMT 可以将 iso-Asp 转化回 Asp，从而修复并有助于细胞存活。本工作描述了使用基因特异性引物从其各自的 cDNA 中扩增出 () 和 () 的 PIMT 基因的分离、克隆、测序和表达，并随后克隆到原核表达载体 pProEXHTa 中。BL21(DE3) 细胞株被用作表达宿主。用不同浓度的 IPTG 和不同时间点研究了两种 PIMTs 的表达动力学。最后，与空载体对照相比，用 PIMT 补充的 BL21(DE3) 细胞被评估了对不同应激的反应。
