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Codon optimization of chicken beta Gallinacin-3 gene results in constitutive expression and enhanced antimicrobial activity in transgenic Medicago sativa L.

文献类型： 外文期刊

作者： Jin, Libo 1 ; Wang, Yunpeng 2 ; Liu, Xiuming 3 ; Peng, Renyi 1 ; Lin, Sue 1 ; Sun, Da 1 ; Ji, Hao 1 ; Wang, Lei 1 ; Zhang, Yuting 1 ; Ahmad, Naveed 5 ;

作者机构： 1.Wenzhou Univ, Inst Life Sci, Wenzhou 325035, Peoples R China

2.Jilin Acad Agr Sci, Changchun 130124, Peoples R China

3.Jilin Agr Univ, Coll Life Sci, Engn Res Ctr Bioreactor & Pharmaceut Dev, Minist Educ, Changchun 130118, Peoples R China

4.Wenzhou Univ, Biomed Collaborat Innovat Ctr Zhejiang Prov, Wenzhou 325035, Peoples R China

5.Shandong Acad Agr Sci, Inst Crop Germplasm Resources, Inst Biotechnol, Shandong Prov Key Lab Crop Genet Improvement, Jinan 250100, Peoples R China

关键词： Chicken ? Gal-3; Medicago sativa L; Antimicrobial activity; Codon optimization

期刊名称：GENE （ 2021影响因子：3.913； 五年影响因子：3.48 ）

ISSN： 0378-1119

年卷期： 2022 年 835 卷

页码：

收录情况： SCI

摘要： Gallinacin-3 (Gal-3) is a newly discovered epithelial beta-defensin that acts as cationic antimicrobial peptides, and plays an important role in chicken innate immunity. However, the gallinacin-3 precursor contained a lengthy C-terminal region, which often hindered its expression. After codon optimization of Gal-3 and construction of an expression vector, the transgenic plants of Medicago sativa were obtained. Transgenic plants were validated and expression of proteins was detected. The antimicrobial activity of chicken ss Gal-3 was analyzed and effects of chicken ss Gal-3 on the body weight and intestinal microflora of mice were described. Our results demonstrated that the codon optimized chicken Gal-3 was stably expressed in transgenic Medicago sativa using the pCAMBIA3301 expression vector under the control of protein phosphatase (Ppha) promoter. Five transgenic plants with the highest expression of chicken ss Gal-3 were selected, and were evaluated for the in vitro antimicrobial activity against Escherichia coli, Staphylococcus aureus and Salmonella typhi. Our findings confirmed that the Minimum Inhibitory Concentration (MIC) of the three bacterial strains were 32, 16 and 128 mu g/mL, respectively. In addition, the effect of chicken Gal-3 on the body weight of mice fed with transgenic plants showed no significant deviation compared with that of the control group. Similarly, no loss of intestinal microflora was evident in the experimental group compared with the control group. Together, our findings demonstrate an alternative method for the stable expression of chicken Gal-3 with significant antibacterial effects and potential probiotics uses. In addition, this study may also be useful in the development of resistant M. sativa plants against pathogenic bacteria in future studies.

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