构建牛CypA基因表达载体并验证其在CHO-K1细胞中的表达
Haidong Liu1, Biyu Zhang1, Meng Zhou1
1College of Animal Science, Anhui Science and Technology University, Chuzhou 233100, China.
Animals : an open access journal from MDPI
|February 13, 2026
概括
研究人员开发了一种新方法,用于生产用于乳腺炎研究的牛类环菲林A (CypA) 蛋白. 这种在CHO-K1细胞中的工程表达载体使得高纯度重组蛋白质的产生成为可能,这对于研究牛的炎症反应至关重要.
科学领域:
- 分子生物学分子生物学
- 生物技术是生物技术.
- 兽医医学 兽医医学 兽医医学
背景情况:
- 在全球范围内,牛乳腺炎是一个重要的经济和动物福利问题.
- 目前的抗生素治疗面临着诸如抗菌素耐药性和药物残留等挑战.
- 环素A (CypA) 是乳腺炎的关键炎症调解剂,需要研究.
研究的目的:
- 构建和验证一个真核细胞表达载体,用于牛类环素A (CypA).
- 建立一个稳定的细胞系来生产重组牛CypA蛋白.
- 促进CypA在牛乳腺炎研究中的体外功能研究.
主要方法:
- 合成并对牛的CypA基因进行了代码优化.
- 将基因克隆到pPB[Exp]矢量中,创建具有EGFP和胺素耐药性的多基斯特朗表达系统.
- 通过限制消化和桑格测序验证了载体的构造;使用qPCR和西欧涂抹在CHO-K1细胞中确认了蛋白质的表达.
主要成果:
- 成功构建了一个牛CypA真核体表达载体,通过测序证实了这一点.
- 在被转染的CHO-K1细胞中证明了EGFP和菌素耐药性的稳定和高效表达.
- 在CHO-K1细胞中通过qPCR和西方 blot成功表达了外源性牛CypA蛋白 (~18kDa).
结论:
- 该研究成功构建并验证了一种牛CypA真核体表达载体.
- 开发的CHO-K1细胞表达系统为重组牛CypA生产提供了一个可靠的平台.
- 这一系统为进一步研究CypA在牛乳腺炎中的作用和治疗潜力奠定了基础.
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