绒毛白蜡FvSAP1基因的克隆与分析及表达载体的构建  

Cloning and Analysis of FvSAP1 Gene in Fraxinus Velutina FvSAP1 Gene and Construction of Expression Vector

作　　者：李丽[1] 臧真荣[1] 燕丽萍[1] 王因花[1] 吴德军[1] LI Li;ZANG Zhengrong;YAN Liping;WANG Yinhua;WU Dejun(Shandong Academy of Forestry/Shandong Provincial Key Laboretory of Forest Tree Genetic Improvement,Shandong Jinan 250014)

机构地区：[1]山东省林业科学研究院山东省林木遗传改良重点实验室,山东济南250014

出　　处：《山东林业科技》2021年第5期7-10,28,共5页Journal of Shandong Forestry Science and Technology

基　　金：山东省重点研发计划项目(2019GNC106143);山东省良种工程项目(2019LZGC009)。

摘　　要：以绒毛白蜡幼苗为试材,Illumina HiSeq 2000高通量测序技术获得的绒毛白蜡转录组Unigene数据为基础,采用RT-PCR方法克隆绒毛白蜡A20/AN1型锌指蛋白基因FvSAP1编码区序列,并通过双酶切等技术构建pCAMBIA2300-FvSAP1植物表达载体。结果表明,从绒毛白蜡中成功克隆出1个A20/AN1型锌指蛋白基因FvSAP1的编码区序列,且成功构建了pCAMBIA2300-FvSAP1植物表达载体。基因编码区长度为846bp,可编码281个氨基酸,分子量为31.0kDa,等电点为8.34,原子总数为4269个,分子式为C1332H2106N400O407S24。蛋白质序列分析表明,FvSAP1具有两个AN1锌指结构,其序列模式为C-X4-C-X(9-12)-C-X(1-2)-C-X4-C-X2-H-X5-H-X-C。Blast分析表明FvSAP1与芝麻(Sesamum indicum,XP_011075237.1)SiSAP16序列的相似性最高,序列一致性为78%。将FvSAP1与同类型的锌指蛋白进行多重序列比较,结果显示该类蛋白序列在不同物种之间具有较高的保守性。Based on the UNIGENE data obtained by high-throughput sequencing technique with Illumina Hiseq 2000,the FvSAP1 coding region of the zinc finger protein gene of Fraxinus Velutina A20/AN1 was cloned by RT-PCR,the pCAMBIA2300-FvSAP1 plant expression vector was constructed by double enzyme digestion.The results shoued that a sequence of zinc finger protein gene FvSAP1 of A20/AN1 type was successfully cloned from Fraxinus Velutina and a plant expression vector pcambia2300-FvSAP1 was constructed.The length of coding region is 846 BP,281 amino acids can be encoded,the molecular weight is 31.0 KDA,the isoelectric point is 8.34,the total number of atoms is 4269,the molecular formula is C1332 H2106 N4000407 S24.Biomolecular structure analysis showed that FvSAP1 had two AN1 zinc finger structures,and its sequence pattern was C-X4-C-X(9-12)-c-x(1-2)-C-X4-C-X2-H-X5-H-X-C.Blast analysis showed that FvSAP1 and Sesamum indicum(XP) Sisap16 had the highest similarity(78%).The result of multiple sequences comparsion between FvSAP1 and the same type of zinc finger proteins showed that the sequence of FvSAP1 was highly conserved among different species.

关 键 词：绒毛白蜡 锌指蛋白 FvSAP1 克隆 植物表达载体 

分 类 号：Q785[生物学—分子生物学]