黄瓜芽黄突变相关基因的初步定位研究-生物化学与分子生物学专业毕业论文.docxVIP

黄瓜芽黄突变相关基因的初步定位研究 黄瓜芽黄突变相关基因的初步定位研究 II II （3）荧光原位杂交过程中的制片的酶解、固定、脱水、探针的长度及标记以及染色等 多个因素会严重影响杂交结果。 关键词：黄瓜 芽黄 hemA1 荧光原位杂交 III III Location of the gene related of the virescent mutation on chromosome of cucumber root tip Author: Guo Hongmei Supervisor:professor GaoJianfeng Abstract Aim:Using the root of cucumber as the material, to obtain the chromosome with the clear and stable C-bands and G-bands ,to differentiate the chromosome better , then located the hemA1 gene encoding the glutamyl-tRNA reductase. Methord: (1) using the improved methord based on the conventional sheeting method， disassociating cell wall hypoosmotic method for observation of chromosome C-bands in cucumber, and obtained the chromosome G-bands by ASG. To analysis the differential gene expression of HemA1 between normal leaves and virescent mutant leaves using real time and polymerase chain reaction detection. locating HemA1 on the chromosome of cucumber by using fluorescencs in situ hybridization. Result: (1)The more clear and stable chromosome C-bands imagine were obtaind by using the improved method than the conventional sheeting method and disassociating cell wall hypoosmotic method . The number of the band on single time chromosome is more than the other methords. The number obtained by improved methord is 30,but it is twelve by the others. The clear G-bands were obtained. The mumber of the G-bands on single time chromosome is 22. the expression amount of HemA1 on the normol leaves of cucumber is more than the expresstion amount on the virescent mutant leaves. The relatively amount is 1.0×103 and 0.38×103 separately. The hybridization signal is weak.The backgroud colour is too deep to examine the hybidization signal. Conclusion:The chromosomes can stable on the different phase pretreated by different reagent. The steps of dissociation,sheeting and drying have great effect on chromosome bands. There aren’t distinct bou