A In situ hybridization experiment system for the floral buds of lilium longiflorum | 麝香百合花蕾组织原位杂交体系的建立
2011
Xi Mengli, Nanjing Forestry University,Nanjing(China) | Pan Mei, Nanjing Forestry University,Nanjing(China) | Shi Jisen, Nanjing Forestry University,Nanjing(China)
chinois. 摘 要：本研究以麝香百合未开放的花蕾为实验材料,以百合花发育相关的B功能基因LLGLO1的特异片段为探针,研究了花蕾切离时间对RNA结构完整性的影响,探讨了蛋白酶K消化浓度及时间对材料内蛋白质消化及材料结构完整性的影响。研究结果表明：花蕾离体1.5 h内RNA仍然保存完整,花蕾离体2 h RNA开始降解,因此,花蕾取材后应在1.5 h之内完成固定。10μg/mL的蛋白酶K在37℃孵育30 min可消化百合花蕾切片中的大部分蛋白质且材料结构完整。原位杂交结果显示LLGLO1基因在四轮花器官中均有表达。
Afficher plus [+] Moins [-]anglais. Abstract Using the floral buds of Lilium longiflorum as materials and the special sequence of LLGLO1 gene as probe,we studied the effect of sampling time on the RNA structure of the floral buds.We observed the influence of proteinase K concentration and time on protein and structural integrity of tissue structure.The results indicated that RNA of 1.5 h sample remained intact,but RNA of 2 h sample began to degrade.Therefore,samples should be fixed the within 1.5 h.Proteinase K of 10μg/mL can be used to incubate for 30 min at 37℃to digest most of the protein of the floral buds.The in situ hybridization result indicated that LLGLO1 gene was expressed in four whorls of floral-organ in Lilium longiflorum.
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