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利用RACE、RT-PCR以及基因组步移技术克隆得到光皮桦Bl COL13全长c DNA(Gen Bank登录号:KP663425)、全长DNA及其启动子序列。该基因DNA全长2 808 bp,具有4个外显子,3个内含子;c DNA全长1 370 bp,包含1 140 bp的开放阅读框,编码含379个氨基酸的蛋白。Bl COL13蛋白N、C端分别有两个B-box和一个CCT结构域,属于典型的BBX蛋白。以拟南芥中BBX蛋白的分类为参照,通过聚类分析表明Bl COL13属于Ⅱ型BBX蛋白;克隆得到的Bl COL13启动子序列长685 bp,含有多个与成花、光响应、激素信号转导以及逆境胁迫相关的顺式作用元件。亚细胞定位试验结果表明该基因主要在细胞核中表达。不同组织中该基因都有一定程度的表达,但在雄花中表达量最高。低温、干旱、ABA和盐胁迫等非生物逆境都在一定程度上诱导Bl COL13的表达,盐胁迫的影响最为明显,表明该基因在光皮桦逆境响应中可能发挥比较重要的作用。
The full length cDNA and its promoter of Bl COL13 were cloned by RACE, RT-PCR and genome walking techniques. The full-length DNA of this gene is 2 808 bp in length with 4 exons and 3 introns. The cDNA is 1 370 bp in length and contains an open reading frame of 1 140 bp and encodes a protein of 379 amino acids. There are two B-boxes and one CCT domain in the N-terminal and C-terminal of COL13 protein, which belongs to the typical BBX protein. Based on the classification of BBX protein in Arabidopsis thaliana, BL COL13 belonged to type II BBX protein by cluster analysis. The BL 131 promoter sequence was 685 bp in length, Guide and cis-acting elements associated with stressors. Subcellular localization test results show that the gene is mainly expressed in the nucleus. The expression of this gene is different in different tissues, but it is the highest in male flowers. Abiotic stress such as low temperature, drought, ABA and salt stress induced the expression of Bl COL13 to a certain extent. The effect of salt stress was the most obvious, indicating that this gene may play an important role in the adversity of Betula luminifera.