Smitha P K, Bathula Christopher, Kumar Anil M, Chandrashekara K N, Dhar Sujan K, Das Manjula
Tumor Immunology Program, MSMF, Mazumdar Shaw Medical Centre, Narayana Health City, 8th floor, Bommasandra, Bangalore, 560 099 Karnataka India.
Department of Biotechnology, Research and Development Centre, Bharathiar University, Coimbatore, 641 046 Tamil Nadu India.
3 Biotech. 2021 Jun;11(6):289. doi: 10.1007/s13205-021-02828-2. Epub 2021 May 22.
Transcription and translation in eukaryotes are distinct processes of the molecular cascade leading to protein production from genetic material. However, establishing correlation between mRNA expression and protein abundance, the end results of the two processes of central dogma, remains a challenge. For transgenic plants, such correlation between mRNA and protein expression serves as a guide to design the transgene, in particular the choices of promoter and codon usage to ensure stable expression of the target protein in relevant tissues under various stress conditions. To elucidate level of mRNA-protein correlation in a commercial transgenic cotton plant , Bollgard II (MON15985), we present the results of Cry1Ac protein expression correlating with corresponding mRNA levels. Protein was quantitated using a home-grown validated ELISA assay with a monoclonal-polyclonal antibody pair, whereas mRNA level was detected by a real-time quantitative PCR assay using standardized reference genes. Our results indicate that protein and mRNA levels are highly correlated in the leaves, but not in squares and stem. The correlations seem to be consistent between young and mature leaves and increase over time of harvesting of samples from months 1-3. These findings demonstrate that transcript level measurement could serve as a proxy to protein abundance for this commercially important cotton species, particularly for leaf tissues which are the most vulnerable organs to cotton bollworms and other pathogens.
The online version contains supplementary material available at 10.1007/s13205-021-02828-2.
真核生物中的转录和翻译是导致从遗传物质产生蛋白质的分子级联的不同过程。然而，在mRNA表达与蛋白质丰度（中心法则这两个过程的最终结果）之间建立相关性仍然是一个挑战。对于转基因植物，mRNA与蛋白质表达之间的这种相关性可作为设计转基因的指南，特别是启动子和密码子使用的选择，以确保目标蛋白质在各种胁迫条件下在相关组织中稳定表达。为了阐明商业转基因棉花品种Bollgard II（MON15985）中mRNA-蛋白质的相关水平，我们展示了Cry1Ac蛋白质表达与相应mRNA水平相关的结果。使用自制的经过验证的ELISA测定法，用单克隆-多克隆抗体对定量蛋白质，而使用标准化参考基因通过实时定量PCR测定法检测mRNA水平。我们的结果表明，蛋白质和mRNA水平在叶片中高度相关，但在棉蕾和茎中不相关。在幼叶和成熟叶之间，这种相关性似乎是一致的，并且在从第1个月到第3个月采集样品的过程中随着时间的推移而增加。这些发现表明，转录水平测量可以作为这种具有商业重要性的棉花品种蛋白质丰度的替代指标，特别是对于叶片组织，叶片组织是棉铃虫和其他病原体最易侵害的器官。
在线版本包含可在10.1007/s13205-021-02828-2获取的补充材料。
