火龙果HuABAR的克隆、生物信息学分析及亚细胞定位
Molecular cloning,bioinformatics analyses and subcelluar localization of HuABAR gene in pitaya (Hylocereus undatus)

在前期利用SSH筛选到抗旱相关基因HuABAR的Unigene序列的基础上，进行了HuABAR的全长cDNA克隆、生物信息学分析及亚细胞定位。结果表明:HuABAR基因cDNA全长为1 239 bp，5′ UTR为264 bp，3′ UTR为414 bp，完整开放阅读框（ORF）共561 bp，编码187个氨基酸；生物信息学分析显示，HuABAR基因编码蛋白具有典型的SRPBCC结构域，并与PYR1/PYLs（pyrabactin resistance 1/PYR1 like）家族具有较高的相似性；HuABAR在干旱、高温（42℃）和低温（4℃）等逆境胁迫下显著上调表达，并在干旱胁迫5 d、高温3 d时表达量最高，低温处理5 d内，表达量持续上升；通过PEG介导法瞬时转化拟南芥原生质体进行亚细胞定位分析，发现该基因定位于细胞质，与已报道的其他PYR1/PYLs家族基因一致。因此，HuABAR基因可能在火龙果干旱胁迫应答中发挥重要作用。
Based on the previous drought associated gene enriched SSH cDNA library and cDNA microarray,we preliminarily screened an HuABAR Unigene presumably involving in the tolerance of abiotic stresses including drought,cold and high temperature.Full length cDNA sequence of this gene was cloned.Bioinformatic analyses and subcellular localization was carried out.The results showed that HuABAR gene was significantly up regulated in response to abiotic stresses including in drought,high temperature and low temperature.The highest expression level was observed at the 5th day or the third day after drought stress or high temperature stress.The expression was increasingly un regulated as exposure to low temperature within five days.HuABAR gene contained 1 239 bp in full cDNA length,and consisted of 264 bp 5′ UTR,414 bp 3′ UTR,and 561 bp open reading frame (ORF) encoding 187 amino acids.This gene might encode a typical SRPBCC domain,and was highly similar to the PYR1/PYLs (pyrabactin resistance 1/PYR1 like) family.The target gene was ligated with green fluorescent protein (GFP) by constructing plant transient expression vector pBWA (V) HS HuABAR gfp,and the PEG mediated method was used to transiently transform into Arabidopsis protoplasts.It was found that target gene was located in the cytoplasm,which satisfied the expectations.It is indicated that HuABAR in pitaya may be involved tolerance of multiple abiotic stresses.