Incorporation of a tag helps to overcome expression variability in a recombinant host

Biotechnology Reports

Asmita Sharda¹  Saikat Bhattacharya¹  Sanjay Gupta¹  Divya Reddy¹  Raja Reddy²  Kakoli Bose² 

[1] Epigenetics and Chromatin Biology Group, Gupta Lab, India;Integrated Biophysics and Structural Biology Lab, Cancer Research Institute, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai 410210, MH, India;

关键词: Rare codons;    Histones;    Truncated GST;    Misfolding;    Translation;   

DOI  :  10.1016/j.btre.2016.06.002

来源: DOAJ

【 摘 要 】

Epigenetics have witnessed a renewed interest over the past decade and assays with recombinant histones has become an important tool for uncovering various aspects of histone biology. However, at times absence of recombinant histone accumulation in bacteria is encountered which is also commonly observed for many eukaryotic proteins in general. In this study, we have investigated the effect of multiple parameters on heterologous expression of proteins. We show that there is marked variability in the accumulation of H2A.2, H2B.1, H3.2 and H4 in the recombinant host, possibly owing to translational variability and degradation by the host proteases. We found that the variability could be overcome by incorporation of the commonly used purification tags, like GST or MBP, of appropriate size and position. Our results provide compelling evidence that transcript parameters like rare codon and GC content, mRNA secondary structure etc. together modulate translation kinetics and govern recombinant protein accumulation.

【 授权许可】

Unknown