Plant Molecular Biology and Genetic Engineering Division, CSIR, National Botanical Research Institute, Rana Pratap Marg, Lucknow 226001, UP, India.
Protein J. 2012 Aug;31(6):477-86. doi: 10.1007/s10930-012-9425-6.
We have optimized and improved the protocol for extraction of histone proteins from Gossypium herbaceum. Histone proteins were isolated by acid extraction method and fractionation of histone proteins were performed using RP-HPLC (reverse-phase high performance liquid chromatography). Analysis of histones from drought tolerant (Vagad) and drought sensitive genotype (RAHS-14) indicated that the tolerant genotype Vagad encodes a 29 kDa protein. Protein sequencing on MALDI TOF/TOF revealed that the 29 kDA protein shared sequence similarity with another drought-inducible linker histone-H1.S reported in tomato. This H1.S like linker histone was not found in RAHS-14 in our study. We further examined the expression of H1 variant at the transcript and protein levels and found that it was induced specifically in the tolerant genotype Vagad.
我们已经优化并改进了从棉草中提取组蛋白的方案。组蛋白通过酸提取法分离，并用反相高效液相色谱（RP-HPLC）进行组蛋白分级。对耐旱（Vagad）和耐旱敏感基因型（RAHS-14）的组蛋白分析表明，耐旱基因型 Vagad 编码一种 29kDa 的蛋白质。MALDI TOF/TOF 上的蛋白质测序表明，29kDa 蛋白与番茄中另一种干旱诱导的连接组蛋白 H1.S 具有序列相似性。在我们的研究中，没有在 RAHS-14 中发现这种 H1.S 样连接组蛋白。我们进一步检查了 H1 变体在转录和蛋白质水平上的表达情况，发现它仅在耐旱基因型 Vagad 中被特异性诱导。
