Morphology and Evolution of Invertebrates, Philipps University Marburg, Marburg, Germany.
Electrophoresis. 2010 Jan;31(2):299-302. doi: 10.1002/elps.200900458.
The model organism Hydra has been used for molecular studies for more than 20 years, however, its DNA base composition has not been determined yet. We have analyzed DNA and total RNA of the freshwater polyp Hydra magnipapillata with two independent procedures of high accuracy and sensitivity - fluorescence labeling of nucleotides followed by CE-LIF detection and (32)P-postlabeling. DNA of Hydra was digested either to deoxyribonucleoside-5'-monophosphates or deoxyribonucleoside-3'-monophosphates selectively conjugated with the fluorescent dye 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionyl ethylene diamine hydrochloride (BODIPY FL EDA) separated and detected using CE-LIF. Both versions of the assay revealed a high A+T composition of 78 and 71%, whereas total DNA methylation (5-methyldeoxycytidine) was 2.6 and 3.1%. Total Hydra RNA showed highest base levels for guanine (33%) and a level of 1.4% for pseudouracil. All values were in good agreement with those determined by the (32)P-postlabeling method.
模式生物水螅已经被用于分子研究超过 20 年，但它的 DNA 碱基组成尚未确定。我们使用两种独立的高精度和高灵敏度方法——荧光标记核苷酸，然后通过 CE-LIF 检测和（32）P 后标记，分析了淡水水螅 Hydra magnipapillata 的 DNA 和总 RNA。水螅的 DNA 分别被消化为脱氧核苷-5'-单磷酸或脱氧核苷-3'-单磷酸，与荧光染料 4,4-二氟-5,7-二甲基-4-硼-3a,4a-二氮杂-s-茚并-3-丙酸乙二胺盐酸盐（BODIPY FL EDA）选择性缀合，然后通过 CE-LIF 分离和检测。两种方法的测定结果均显示出高达 78%和 71%的 A+T 组成，而总 DNA 甲基化（5-甲基脱氧胞嘧啶）水平分别为 2.6%和 3.1%。水螅总 RNA 的鸟嘌呤碱基水平最高（33%），假尿嘧啶水平为 1.4%。所有这些值与（32）P 后标记方法测定的值非常吻合。
