猪干扰素-α基因的克隆及其植物表达载体的构建

Molecular Cloning and Construction of Plant Expressive Vector of Porcine Interferon-α

目的:克隆与分析猪干扰素-α(INF-α)基因,构建猪干扰素基因的高效植物表达载体。方法:根据NCBI中DQ248997序列设计引物,以猪的总DNA为模板,PCR扩增出猪的INF-α基因,克隆至pBS-T载体后进行序列分析,构建猪干扰素基因的植物表达载体。结果:实验所克隆序列经Blastn比对,98%的核酸序列相同,98%的蛋白质序列相同,3个非功能性氨基酸与基因库中序列不一致,推测为猪INF-α的一个亚型。构建的2个植物表达载体经BamHⅠ/SacⅠ限制性内切酶消化,均可得到570bp的目的基因。结论:成功克隆了猪的INF-α基因,并构建出含猪INF-α基因的高效植物表达载体pBI121/INF和pCAMBIA1301/INF。

Objective： To clone and sequence porcine interferon-α（INF-α） gene, construct efficient plant expressive vector of porcine interferon. Methods： Designed primers according to DQ248997 of NCBI. Amplified the target porcine INF-α gene by PCR protocol from the porcine genome DNA, then cloned into pBS-T Vector and sequenced. Constructed plant expressive vector of porcine INF-α. Results： The gene cloned was compared by Blastn, 98% nucleic acids were the same, 98% proteins were the same, three no function amino acids differed from the sequence in GenBank. Confered that it was a subunit model of porcine interferon-α. Both of the two plant expressive vectors could be digested by BamH Ⅰ/Sac Ⅰ restriction enzymes and got the target fragment（570 bp）. Conclusion： Successfully cloned the porcine interferon-α gene, and constructed two efficient plant expressive vectors pBI121/INF and pCAMBIA1301/INF of porcine interferon-α.