棉蚜细胞色素P450 CYP6J1的克隆与表达
发布时间：2019-07-26 05:38
【摘要】：为了深入了解细胞色素P450 CYP6J1蛋白的结构和功能,本实验克隆获得了棉蚜Aphis gossypii P450CYP6J1基因,对该基因进行信息学及原核表达分析,通过SDS-PAGE检测目的蛋白的表达结果,并用Western-blot进行验证。结果表明,P450 CYP6J1序列长1 398 bp,编码氨基酸数为465,理论分子量为53.67 k D,理论等电点为8.80。氨基酸序列分析表明该序列具有完整的开放阅读框,且没有信号肽。同源性分析表明,棉蚜c DNA序列推导的氨基酸与豌豆蚜Acyrthosiphon pisum的保守性最为接近,一致性可达92%。在大肠杆菌Escherichia coli BL21中表达获得的His-CYP6J1蛋白,并用Western-blot检测目的蛋白大小正确。这些研究结果为棉蚜P450 CYP6J1多克隆抗体制备提供了基础。
[Abstract]:In order to understand the structure and function of cytochrome P450CYP6J1 protein, the Aphis gossypii P450CYP6J1 gene of APHIS gossypii was cloned and analyzed by informatics and prokaryotic expression. The expression of the target protein was detected by SDS-PAGE and verified by Western-blot. The results show that the length of P450 CYP6J1 sequence is 1.398 bp, the amino acid number is 465, the theoretical molecular weight is 53.67 KD, and the theoretical isoelectric point is 8.80. Amino acid sequence analysis showed that the sequence had a complete open reading frame and no signal peptide. Homology analysis showed that the amino acids deduced from the c DNA sequence of APHIS gossypii were the closest to the conservation of Acyrthosiphon pisum of pea aphid, and the consistency was 92%. The His-CYP6J1 protein was expressed in E. coli Escherichia coli BL21 and the target protein was detected by Western-blot. These results provide a basis for the preparation of polyclonal antibody against APHIS gossypii P450 CYP6J1.
【作者单位】： 新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室;中国农业大学农学与生物技术学院;
【基金】：国家自然科学基金项目(31330064,31471781)
【分类号】：Q78;S433
，
本文编号：2519367
[Abstract]:In order to understand the structure and function of cytochrome P450CYP6J1 protein, the Aphis gossypii P450CYP6J1 gene of APHIS gossypii was cloned and analyzed by informatics and prokaryotic expression. The expression of the target protein was detected by SDS-PAGE and verified by Western-blot. The results show that the length of P450 CYP6J1 sequence is 1.398 bp, the amino acid number is 465, the theoretical molecular weight is 53.67 KD, and the theoretical isoelectric point is 8.80. Amino acid sequence analysis showed that the sequence had a complete open reading frame and no signal peptide. Homology analysis showed that the amino acids deduced from the c DNA sequence of APHIS gossypii were the closest to the conservation of Acyrthosiphon pisum of pea aphid, and the consistency was 92%. The His-CYP6J1 protein was expressed in E. coli Escherichia coli BL21 and the target protein was detected by Western-blot. These results provide a basis for the preparation of polyclonal antibody against APHIS gossypii P450 CYP6J1.
【作者单位】： 新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室;中国农业大学农学与生物技术学院;
【基金】：国家自然科学基金项目(31330064,31471781)
【分类号】：Q78;S433
，
本文编号：2519367
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