Efficient Seamless Cloning of Full-length and Truncated GFP-tagged FAM210B Plasmids and Comparison with Traditional Restriction Cloning

Donglan Huang ； Xiaofen Zheng ； Qionghua Lian ； Xuejuan Gao ； Qiuyu Liu

《International Core Journal of Engineering》 8卷6期 (2022/06) Pp. 135-142

https://doi.org/10.6919/ICJE.202206_8(6).0020

摘要

The aim was to construct a series of expressing plasmids containing full-length or truncated FAM210B cDNA fused with GFP by seamless cloning technique. According to the design principle of seamless cloning, primers were designed and synthesized to amplify corresponding cDNA. On the other hand, the GFP-vector plasmid was linearized by restriction enzyme digestion, then they were ligated with each other by homologous recombinant kit. Finally, the competent E. coli DH5 was used to transform the linked products. In this study, we successfully constructed the fusion plasimds of FAM210B or its truncates with GFP by seamless cloning technology and verified by sequencing. The expressions of various recombinant GFP-tagged fusion plasimds were confirmed by fluorescence microscopy after transfection into H1299 cells. The successful construction and expression of these fusion plasmids would provide tools for the functional study of FAM210B in the future.

關鍵字

Seamless Cloning ； Restriction Cloning ； FAM210B ； Structural Domain

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