摘要
目的 通过对贝母几个种遗传多态性的研究来开发在分子水平上用于鉴别贝母基因型及不同种类的DNA芯片技术。方法 用PCR扩增法和DNA直接测序法确定核苷酸多态性 ,用DNA芯片进行基因检测。结果 首先提取来自多种贝母根茎的基因组DNA ,对 2 6SrDNA基因D2与D3区的多态性片段进行扩增和测序 ,然后将不同种属多态性片段的特异性寡核苷探针点置于经多聚赖氨酸处理包被的芯片。用来自不同种贝母的荧光素标记的PCR产物与DNA芯片进行杂交 ,可在芯片特定位置检测到不同种贝母的荧光信号。结论 本研究显示DNA芯片技术可为植物种属的验证与质量控制提供一种快速。
Aim To investigate the genetic polymorphism of several species of Fritillaria and to develop a DNA chip for the genotyping and identification of the origin of various species of Fritillaria at molecular level. Methods Genomic DNA from bulbs of several Fritillaria species was extracted and the polymorphisms of the D2 and D3 regions inside the 26S rDNA gene were identified by direct sequencing. Oligonucleotide probes specific for these polymorphisms were designed and printed on the poly lysine coated slides to prepare the DNA chip. PCR products from the Fritillaria species were labeled with fluorescence by incorporation of dye labeled dideoxyribonucleotides and hybridized to the immobilized probes on the chip. Results The polymorphisms were used as markers for discrimination among various species. Specific oligonucleotide probes were designed and immobilized on a DNA chip. Differentiation of the various Fritillaria species was accomplished based on hybridization of fluorescent labeled PCR products with the DNA chip. Conclusion The results demonstrated the reliability of using DNA chips to identify different species of Fritillaria , and the DNA chip technology can provide a rapid, high throughput tool for genotyping and quality assurance of the plant species verification.
出处
《药学学报》
CAS
CSCD
北大核心
2003年第3期185-190,共6页
Acta Pharmaceutica Sinica
基金
CityUniversityofHongKong (960 0 0 0 1)
