【摘要】：為了深入了解細(xì)胞色素P450 CYP6J1蛋白的結(jié)構(gòu)和功能,本實(shí)驗(yàn)克隆獲得了棉蚜Aphis gossypii P450CYP6J1基因,對(duì)該基因進(jìn)行信息學(xué)及原核表達(dá)分析,通過(guò)SDS-PAGE檢測(cè)目的蛋白的表達(dá)結(jié)果,并用Western-blot進(jìn)行驗(yàn)證。結(jié)果表明,P450 CYP6J1序列長(zhǎng)1 398 bp,編碼氨基酸數(shù)為465,理論分子量為53.67 k D,理論等電點(diǎn)為8.80。氨基酸序列分析表明該序列具有完整的開(kāi)放閱讀框,且沒(méi)有信號(hào)肽。同源性分析表明,棉蚜c DNA序列推導(dǎo)的氨基酸與豌豆蚜Acyrthosiphon pisum的保守性最為接近,一致性可達(dá)92%。在大腸桿菌Escherichia coli BL21中表達(dá)獲得的His-CYP6J1蛋白,并用Western-blot檢測(cè)目的蛋白大小正確。這些研究結(jié)果為棉蚜P450 CYP6J1多克隆抗體制備提供了基礎(chǔ)。
[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.
【作者單位】： 新疆大學(xué)生命科學(xué)與技術(shù)學(xué)院新疆生物資源基因工程重點(diǎn)實(shí)驗(yàn)室;中國(guó)農(nóng)業(yè)大學(xué)農(nóng)學(xué)與生物技術(shù)學(xué)院;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(31330064,31471781)
【分類(lèi)號(hào)】：Q78;S433
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本文編號(hào)：2519367