Molecular Biology and Genetic Engineering Laboratory, Defence Institute of Bio-Energy Research (DIBER), DRDO, Goraparao, P.O.-Arjunpur, Haldwani, Nainital, 263139, Uttarakhand, India.
Mol Biol Rep. 2013 Jul;40(7):4235-40. doi: 10.1007/s11033-013-2505-7. Epub 2013 May 4.
Glycerol-3-phosphate acyltransferase (GPAT) catalyzes first and the rate limiting step in glycerolipid synthesis pathway, which in turn contribute to stabilization of plasma membrane structure and oil lipid synthesis in plant cells. Here, we report cloning and characterization of GPAT gene from Lepidium latifolium (LlaGPAT). The cDNA sequence (1,615 bp) of LlaGPAT gene consisted of 1,113 bp ORF encoding a protein of 370 aa residues, with deduced mass of 41.2 kDa and four acyltransferase (AT) motifs having role in catalysis and in glycerol-3-phosphate binding. Southern blot analysis suggested presence of a single copy of the gene in the genome. Tissue specific expression of the gene was seen more abundantly in aerial parts, compared to the roots. Quantitative real-time PCR indicated down-regulation of the gene by cold (4 °C), drought (PEG6000), salt (300 mM NaCl) and ABA (100 μM) treatments. Considering the vitality of the function of encoded enzyme, LlaGPAT can be considered a potential candidate gene for genetic engineering of oil yields and abiotic stress management in food as well as fuel crops.
甘油-3-磷酸酰基转移酶（GPAT）催化甘油脂质合成途径中的第一步和限速步骤，这反过来又有助于稳定植物细胞的质膜结构和油脂合成。在这里，我们报告了来自宽叶独行菜（Lepidium latifolium）的 GPAT 基因的克隆和特性。LlaGPAT 基因的 cDNA 序列（1615bp）由 1113bp 的 ORF 组成，编码 370 个氨基酸残基的蛋白质，推断分子量为 41.2kDa，有四个酰基转移酶（AT）基序在催化和甘油-3-磷酸结合中起作用。Southern blot 分析表明该基因在基因组中存在一个拷贝。与根部相比，该基因在地上部分的组织特异性表达更为丰富。定量实时 PCR 表明，低温（4°C）、干旱（PEG6000）、盐（300mM NaCl）和 ABA（100μM）处理均导致该基因下调。考虑到编码酶的活力，LlaGPAT 可以被视为提高油料作物产油量和非生物胁迫管理的遗传工程的潜在候选基因，适用于食品和燃料作物。
