Secretory expression of ribosomal protein S11 in Pichia pastoris and its therapeu-tic potential in melanoma
10.16303/j.cnki.1005-4545.2025.08.14
- VernacularTitle:核糖体蛋白S11在毕赤酵母中的分泌表达及其对黑色素瘤的治疗作用
- Author:
Xiaomei ZHANG
1
;
Jiangang ZHANG
1
;
Chengkai ZHOU
1
;
Qi YIN
1
;
Zhenzhen LIU
1
;
Yongjun YANG
1
;
Wei CHEN
1
Author Information
1. 吉林大学 动物医学学院,吉林 长春 130062
- Publication Type:Journal Article
- Keywords:
Pichia pastoris;
ribosomal protein S11;
trained immunity;
tumor
- From:
Chinese Journal of Veterinary Science
2025;45(8):1688-1695
- CountryChina
- Language:Chinese
-
Abstract:
The ribosomal protein S11(RPS11)from Enterococcus faecalis is was heterologously ex-pressed using the Pichia pastoris system.The RPS11 gene sequence was optimized to match the yeast codon preference,and the recombinant expression vector pPIC9K-RPS11 was constructed.Electroporation was used to transform the vector into the Pichia pastoris GS115 strain,and high-copy recombinant strains were selected through G418 resistance screening.Protein expression was induced with methanol,and the expression was verified by SDS-PAGE.The recombinant protein was applied in a mouse melanoma treatment model to evaluate its therapeutic effects.The results showed that the recombinant expression vector pPIC9K-RPS11 successfully expressed the target protein with an approximate molecular weight of 14 kDa in Pichia pastoris.The optimal fermenta-tion conditions were determined to be an induction temperature of 30 ℃,induction time of 72 h,and methanol concentration of 1%.Analysis using a mouse peritoneal macrophage trained immuni-ty model revealed that recombinant RPS11 possessed biological activity capable of inducing trained immunity.Additionally,therapeutic experiments in a mouse melanoma model demonstrated that recombinant RPS11 significantly inhibited tumor growth.These findings suggest that the recombi-nant RPS11 secreted by Pichia pastoris not only possesses biological activity in inducing trained immunity but also inhibits tumor cell growth in a mouse melanoma model,providing theoretical support for the heterologous expression and potential applications of recombinant RPS11.