CAJ | 학술논문

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26S rDNA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、高通量的检测工具.
목적통과대패모궤개충유전다태성적연구래개발재분자수평상용우감별패모기인형급불동충류적DNA심편기술.방법용PCR확증법화DNA직접측서법학정핵감산다태성,용DNA심편진행기인검측.결과수선제취래자다충패모근경적기인조DNA,대26S rDNA기인D2여D3구적다태성편단진행확증화측서,연후장불동충속다태성편단적특이성과핵감탐침점치우경다취뢰안산처리포피적심편.용래자불동충패모적형광소표기적PCR산물여DNA심편진행잡교,가재심편특정위치검측도불동충패모적형광신호.결론본연구현시DNA심편기술가위식물충속적험증여질량공제제공일충쾌속、고통량적검측공구.
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.