Ultraviolet (UV) radiation could induce pyrimidine-related dimeric lesions in genomic DNA. Though the cyclobutane pyrimidine dimers (CPDs) are the most abundant UV-induced lesions, the pyrimidine (6-4) pyrimidone photoproducts (6-4PPs) may have more serious, potentially lethal, and mutagenic effects. It is important to have 6-4PP-containing oligodeoxynucleotides to be prepared for studying their adverse biological effects. Here, we developed a UV-irradiated water droplet method for the preparation of a biotinylated, 6-4PP-containing 10-mer oligodeoxynucleotide. By the use of HPLC purification and enrichment twice, the final yield is estimated to be about 8.1%. In contrast, without applying droplet technique, the direct UV irradiation against oligonucleotide-containing aqueous solution, the product yield is very low. The enzymatic hydrolyzation of the obtained product shows a 6-4PP characteristic ion transition of 545.12 → 432.13 in negative ion mode UHPLC-Q-TOF/MS. The established procedure for the preparation of 6-4PP-containing oligonucleotides is convenient with an improved yield.

Open image in new window

紫外线（UV）可以在基因组DNA中诱导嘧啶相关的二聚体损伤。尽管环丁烷嘧啶二聚体（CPD）是紫外线引起的最丰富的病变，但嘧啶（6-4）嘧啶酮光产物（6-4PPs）可能具有更严重的，潜在的致死性和诱变作用。重要的是要制备含6-4PP的寡脱氧核苷酸，以研究其不利的生物学作用。在这里，我们开发了一种紫外线照射的水滴法，用于制备生物素化的，含6-4PP的10-mer寡脱氧核苷酸。通过使用HPLC纯化和富集两次，最终产率估计为约8.1％。相反，如果不使用液滴技术，直接对含有寡核苷酸的水溶液进行紫外线照射，则产品收率非常低。所得产物的酶促水解在负离子模式UHPLC-Q-TOF / MS中显示545.12→432.13的6-4PP特征离子跃迁。用于制备含6-4PP的寡核苷酸的既定程序方便且产率提高。

在新窗口中打开图像