【作者】 张雅娟；

【导师】 蒋滢； 王尉平；

【作者基本信息】 苏州大学 ， 生物化学与分子生物学， 2001， 硕士

【摘要】 本研究以11组随机引物（P₁、P₂、P₃、P₄、P₅、P₆、P₇、P₈、P₉、K、W）对不同产浆量的四品系西蜂（低产浆：喀尼阿兰蜂；高产浆：浙农大、平湖及肖山等意蜂）的基因组DNA进行RAPD—PCR遗传分析。以W（5’-CGGCCCCGGT-3’）为随机引物通过RAPD—PCR扩增技术从西蜂的不同三品系（浙农大、平湖和肖山等意蜂）的DNA分子中获得了与高产浆相关的共同特异基因标记—W316bp片段，用非放射性标记物—地高辛（DIG），使用随机引物标记法，制备成DIG-W316bp探针，再与高、低产浆西蜂的基因组DNA及PCR扩增产物分别进行斑点杂交及Southern杂交鉴定。结果：（1）以W为引物，在高产王浆的浙农大、平湖、肖山等三品系意蜂的基因组DNA的扩增产物中找到了与高产浆优良性状相关的特异基因标记——W316bp片段。（2）然后将W316标记成DIG-W316bp探针，再与高、低产王浆西蜂的DNA基因组及扩增产物分别进行斑点杂交、Southern杂交以此鉴别高、低产王浆西蜂。（3）发现W316bp探针只与高产王浆西蜂的PCR扩增产物及其基因组DNA杂交，而不与低产浆西蜂的PCR扩增产物及其基因组DNA杂交。以上结 从西羹DNA分子中寻找与高产王浆用关的特异基因标记及其鉴定 摘要 果提示：W3 16hP对产浆高的西蜂（浙农大、平湖和肖山等意蜂）是特必 异的，W3 6hP与产浆高西蜂基因组是同源的，由此可以推断W316bP 可能是高产浆西蜂优良性状相关的特异基因标记。应用 W3 6hP探针对 产浆量不同的西蜂可以在实验室进行快速N8h)鉴别，确定高产浆蜂 群，为蜂农在分子水平上选择高产浆的西蜂提供了科学依据，同时为 进一步寻找高产浆西蜂的相关基因打下了良好基础。更多还原

【Abstract】 In this report, 11 random primers (P1, P2, P3, P4, P5, P6, P7, P8, P9, K, W) and RAPD (Random Amplified Polymorphic DNA)-PCR were used to amplify and hereditarily analyze the genomic DNA in four race of Apis mellifera Lindauer (A.m.Lindauer of low royal jelly quantity-carnica; A.m.Lindauer of high royal jelly quantity: Zhenongda , Pinghu and Xiaoshan of A.m.ligustica) which were different in royal jelly quantity. W316bp (bp, base pair) is a common special DNA fragment used PCR with random primer W (5’-CGGCCCCGGT-3’) in 3 race of Apis mellifera Lindauer (Zhenongda, Pinghu and Xiaoshan of high royal jelly quantity). W316bp was marked with digoxigenin (DIG). Probe DIG-W316bp hybridized with PCR products and genomic DNA of A.m.Lindauer of high and low royal jelly quantity.The results showed :(1) Primer W produced a special W316bp fragment of DNA which was found in Zhenongda, Pinghu and Xiaoshan of high royal jelly quantity. (2)W316bp labelled with digoxigenin (DIG) may be used to identify special fragment of high or low royal jelly quantity-hybridized by Dot blotting with genomic DNA and Southern blotting with PCR products of A.m.Lindauer. (3) Probe DIG-W316bp only hybridized with 3 races of A.m.Lindauer of high royal jelly quantity and didn’t hybridize with A.m.Lindauer of low royal jelly quantity. Bees of high royal jelly quantity appeared the positive signal.The results suggest: It is important that A.m.Lindauer of high royal jelly quantity has the special gene marker-W316bp. It is proved that W316bp is a special DNA fragment in A.m.Lindauer of high royal jelly quantity. W316bp is related to the trait of high royal jelly quantity in A.m.Lindauer. W316bp may be useful to look for the gene, which is related to high royal jelly quantity in A.m.Lindauer.Postgraduate : Zhang Yang-juan Directed by Professor Jiang YingAssociate Professor Wang Wei-ping更多还原