甘蓝型油菜LPAT5基因克隆与表达分析

Cloning and Expression Analysis of LPAT5 from Brassica napus

克隆并对溶血磷脂酸酰基转移酶时空表达和逆境表达进行分析,旨在为进一步研究其生物学功能奠定基础。结合同源克隆与半定量RT-PCR的方法,克隆得到甘蓝型油菜湘油15 LPAT5的1条全长1 041 bp的CDs序列,并命名为BnLPAT5。序列分析表明,它具有LPLAT_LCLAT1样结构域,属于LPLAT超基因家族。时空表达分析表明,BnLPAT5在根中的表达量最高,是茎和胚的2倍。逆境分析表明,BnLPAT5在NaCl、PEG4000、水渍、6BA和ABA的胁迫下呈现2种不同的表达模式,他们均对BnLPAT5基因的表达具有抑制作用。在NaCl、PEG4000及水渍处理下,BnLPAT5呈"骤降缓升"的表达模式,处理3 h时,达到最低表达量。对于6BA和ABA的处理,BnLPAT5却呈现"缓降缓升"的表达模式,处理12 h时,表达量最低。极差分析显示,高盐、干旱、水渍、6BA和ABA对BnLPA5基因的表达影响程度相当,相对而言,高盐对其表达的影响较大。试验首次从甘蓝型油菜中克隆得到LPAT5的1个拷贝,并分析了其时空表达和逆境表达,认为所得基因为普遍表达基因,并对干旱比较敏感。

In order to further study on biological function of BnLPAT5,we cloned and analysed this gene for its temporal and spatial expression and stress expression. Combined with the technology of homology-based cloning and semi-quantitative RT-PCR,one copy of LPAT5 full-length CDs sequence（ 1 041 bp） had been cloned from Brassica napus of Xiangyou 15 in our study,and designated as BnLPAT5. Sequence analysis revealed that BnLPAT5 had the LPLAT_ LCLAT1 like domain and belonged to the LPLAT superfamily. Temporal and spatial expression results showed that the highest expression of BnLPAT5 was in root and the expression levels was two times of stem and embryo. Stress analysis indicated that BnLPAT5 presented two kinds of expression patterns under the treatments of NaCl,PEG4000,waterlogging,6BA and ABA,and which limited to the expression of BnLPAT5. Under the treatment of NaCl,PEG4000 and water logging,BnLPAT5 presented a model of,rapidly fall and then slowly ascent and the minimum was at 3 h after treatment. As for the treatment of 6BA and ABA,BnLPAT5 presented a model of slowly ascent and then slowly fall＇ and the minimum was at 12 h after treatment. Compaired the influence of,high-salt,arid,water logging,6BA and ABA,range analysis showed those factors regulated the expression of BnLPAT5 in the similarity level. In comparison,high-salt＇s inflluence was more obvious. A copy of LPAT5 was isolated from Brassica napus and analysed its temporal and spatial expression and stress expression for the first time. We thought BnLPAT5 was a ubiquitous expression gene and sensitive to the arid.