【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-FvSA P1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FvSA P1的编码区序列,且成功构建了pCAMBIA2300-FvSA P1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FvSA P1与芝麻（Sesamum indicum,XP₀11075237.1）SiSAP 16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。更多还原

【Abstract】 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.更多还原