扬州大学教师个人主页服务平台袁玉国--中文主页-- The goat β‑casein/CMV chimeric promoter drives the expression of hLF in transgenic goats produced by cell transgene microinjection

袁玉国

学位：博士
职称：副高级
所在单位：兽医学院

博士生导师

硕士生导师

教师英文名称：Yuguo Yuan
教师拼音名称：Yuan Yuguo
电子邮箱：
入职时间：2010-06-01
学历：博士研究生毕业
办公地点：扬州大学文汇路校区45楼102室
性别：男
联系方式：0514-87979228
在职信息：在岗
毕业院校：扬州大学

学科：临床兽医学

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The goat β‑casein/CMV chimeric promoter drives the expression of hLF in transgenic goats produced by cell transgene microinjection

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发表刊物：

Int J Mol Med

摘要：

There is growing interest in the application of lactoferrin (LF) as a drug or food additive for animals and humans. The objective of this study was to produce transgenic cloned goats that would serve as living bioreactors, expressing high levels of recombinant human LF (rhLF) in their milk. We designed a pCL25 expression vector containing goat β‑casein/CMV chimeric promoter in order to facilitate rhLF expression. This pCL25‑rhLF‑Neo vector was microinjected into goat fetal fibroblasts. G418 selection and PCR analysis were used to identify transgenic donor cells suitable for somatic cell nuclear transfer (SCNT). After SCNT and embryo transplantation, goats harboring the hLF gene were produced, as confirmed via PCR and southern blotting. The average rhLF concentration in milk from this transgenic goat was 3.89 mg/ml as determined via ELISA. We also used an optimized buffer in order to effectively elute high‑purity (95.8%) rhLF from a cation‑exchange column, with the recovered rhLF exhibiting high biological activity. Findings from this study demonstrated that it is possible to generate a transgenic goat harboring the hLF transgene driven by the goat β‑casein/CMV chimeric promoter. It represents an initial step towards the production of rhLF, potentially allowing for industrialized purification in the future.

论文类型：

Research Atricle

是否译文：

否

发表时间：

2019-12-05

收录刊物：

SCI

发布期刊链接：

https://pubmed.ncbi.nlm.nih.gov/31661123/

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下一条： Accurately cleavable goat β-lactoglobulin signal peptide efficiently guided translation of a recombinant human plasminogen activator in transgenic rabbit mammary gland

个人简介

袁玉国，博士，副教授，硕士、博士生导师。2010年毕业于扬州大学兽医学院，获临床兽医学博士学位，并留校任教。2015.10—2017.7韩国Konkuk University博士后，2017.9—2018.9韩国Gyeongsang National University访问学者。

长期从事动物体细胞克隆和转基因克隆的研究工作，2007年在国内外首次报道利用成年乳腺上皮细胞获得克隆山羊，先后获得转人乳铁蛋白，白蛋白等转基因克隆羊。利用TALEN，Cas9技术获得Fel d1基因敲除猫、MSTN基因敲除山羊，人高血脂兔动物模型（兔）等。同时开展了功能纳米材料的制备及其在抗菌、抗肿瘤和化妆品上的应用。先后主持了国家自然科学基金、国家转基因重大专项子课题、江苏省种业“揭榜挂帅”等15余项课题的研究工作，在 EBioMedicine，Int J Nanomedicine，Cells，Oxid Med Cell Longev, Int J Mol Sci，Mol Reprod Dev，Theriogenology等杂志发表SCI论文30余篇，获得省部级等科技奖励2项、发明专利1项。

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