当前位置: 首页 > 文章 > 衣藻肌动蛋白基因的克隆及核苷酸序列分析 中国农业大学学报 1993 (S1) 19-26
Position: Home > Articles > Cloning and Sequencing of an Actin Gene from Chlamydomonas reinhardtii Journal of China Agricultural University 1993 (S1) 19-26

衣藻肌动蛋白基因的克隆及核苷酸序列分析

作  者:
王荣臣;胡松年;祁岩;郭静成;阎隆飞
单  位:
北京农业大学生物学院生物化学研究室
关键词:
衣藻;Chlamydomonas reinhardtii;肌动蛋白基因;内含子;拷贝数
摘  要:
提取衣藻总 DNA,以此为模板并参照肌动蛋白基因编码区端的序列合成寡聚核苷酸引物,进行聚合酶链式反应,扩增到一个1.2 kb 的 DNA 片段.将此片段进行克隆并经分子探针杂交,证明此片段为衣藻肌动蛋白基因.经限制性核酸内切酶处理,构建了衣藻肌动蛋白基因的物理图谱.根据物理图谱进行亚克隆以后,测得了编码区5′端的618个核苷酸的顺序.衣藻肌动蛋白基因的编码序列与高等植物之间的同源性大于90%,高于与高等动物、原生动物及真菌的同源性.但在基因的结构上,衣藻肌动蛋白基因又明显地不同于高等植物,在已经测定的基因片段上,没有发现内含子的存在.经限制性内切酶片段多态性分析,衣藻中含有一个肌动蛋白基因拷贝.
译  名:
Cloning and Sequencing of an Actin Gene from Chlamydomonas reinhardtii
作  者:
Wang Rongchen Hu Songnian Qi Yan Guo Jingcheng Yan Longfei (College of Biology,Beijing Agricultural University,Beijing 100094)
关键词:
Chlamydomonas reinhardtii;;actin gene;;intron;;copy number
摘  要:
Total DNA was isolated from Chlamydomonas reinhardtii ,a unicellular alga.The protein coding region of actin gene was amplified by polymerase chain reaction from this DNA preparation.The amplified DNA fragment of approximately 1.2 kb was inserted into the plasmid pBluescript and then the re-combinant plasmid was transformed into Escherichia coil Actin gene clones were identified by Southern hybridization with an actin cDNA clone from pea tendrils as the probe. The nucleotide sequence of the DNA clone was partially determined.The sequenced region of 618 bp had a homology of 80% with the corresponding sequence of pea actin cDNA PEAcl.The deduced amino acid sequence of this fragment was 94% homologous to the counter part of PEAcl.These are much higher than the homologies between actin gene of Chlamydomonas and those of animals and lower eukaryotic or- ganisms.RFLP analysis showed that C.reinhardtii had one copy of act in gene.

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