Barnett R I, Gray V A, MacKinnon E A
Histochemistry. 1980;65(3):207-15. doi: 10.1007/BF00493170.
In an effort to minimize subjective bias, a classification scheme was devised to assess Giemsa staining patterns obtained with experiments involving acetic acid-alcohol and exogenously applied histone 1 and polypeptides. A single rinse of metaphase preparations with acetic acid-alcohol quantitatively reduced Giemsa dye binding. Acid-alcohol irreversibly changed the conformation of H1 and its ability to interfere with trypsin G-banding. Our results suggest that, in addition to protein extraction, acid-alcohol may alter the conformation of acid-insoluble components of metaphase chromosomes. The carboxy-terminal polypeptide (residues 73--212) from NBS cleavage of H1 was an effective inhibitor of Giemsa staining and trypsin G-banding. However, this polypeptide which is preferential for supercoiled DNA was much less efficient in inhibiting Giemsa staining of trypsinized metaphase chromosomes. The molecular consequences of these experiments are discussed.
为尽量减少主观偏差，设计了一种分类方案，以评估在涉及醋酸 - 酒精以及外源性应用组蛋白1和多肽的实验中获得的吉姆萨染色模式。用醋酸 - 酒精对中期染色体标本进行单次冲洗可定量减少吉姆萨染料结合。酸 - 酒精不可逆地改变了H1的构象及其干扰胰蛋白酶G显带的能力。我们的结果表明，除了蛋白质提取外，酸 - 酒精可能会改变中期染色体酸不溶性成分的构象。H1经NBS裂解产生的羧基末端多肽（第73 - 212位氨基酸残基）是吉姆萨染色和胰蛋白酶G显带的有效抑制剂。然而，这种对超螺旋DNA具有偏好性的多肽在抑制胰蛋白酶处理后的中期染色体的吉姆萨染色方面效率要低得多。讨论了这些实验的分子结果。
