人工合成的简单重复序列的克隆及其应用  

Cloning of Synthetic Simple Repetitive Sequences and Their Uses in DNA Fingerprinting of Animals

作　　者：范青林 孟安明[1] 周顺伍[1] 齐顺章[1] 

机构地区：[1]北京农业大学生物学院动物生化室,北京100094

出　　处：《农业生物技术学报》1994年第2期81-87,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金

摘　　要：人工合成的简单重复序列（TG）_7/（AC）_7、（GTG）_5/（ACC）_5和（GAG）_5/（TCC）_5分别连接成多聚体（TG）_n/（CA）_n、（GTG）_n/（CAC）_n和（GAG）_n（CTC）_n,回收大于300bp的多聚体直接进行放射性标记做为DNA指纹探针,在鸡、鸭、鹌鹑、天鹅、猪、牛、鼠上产生具有高度变异性的DNA指纹图。回收的大于300bp多聚体还被克隆到质粒pGEM-3Zf（一）中。对重组质粒进行的酶切和序列分析表明,这些高度重复序列在扩增过程中不稳定,发生了序列的缺失和重排。尽管如此,这些重组质粒仍可用做DNA指放探针,在有关动物上产生个体特异性的DNA指纹图。与用未克隆的多聚寡核苷酸探针产生的图谱相比,经克隆的相应多聚寡核苷酸探针产生的图谱信号更强,但带型未变。Synthetic simple repetitive sequences (TG)7/(AC)7, (GTG)5/(ACC)5 and (GAG)5/(ACC)5 were ligated to form polymers of (TG)n/(CA)n,(GTG)5/(CAC)n and (GAG)n/(CTC)n, respectively. The polymers of greater than 300 bp were recovered and radiolabeled as DNA fingerprinting probes. They generated highly variable DNA fingerprints in chicken, duck, quail, swan, pig, cattle and mouse. The bigger polymers were cloned into Smal site of plasmid pGEM-3zf (-). Restriction and sequence analyses revealed that the cloned highly repetitive sequences underwent deletions and rearrangements during propagation in E. clot. Nevertheless, these recombinant plasmids could be used as DNA fingerprinting probes and produced individual-specific DNA fingerprints plasmids in several animal species. Compared to the uncloned polymers, as hybridization probes, the cloned counterparts gave higher hybridization signals without changing hybridization patterns.

关 键 词：简单重复序列 DNA 克隆 动物 

分 类 号：S188[农业科学—农业基础科学] Q347[生物学—遗传学]