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耳鲍(Haliotis asinina)核糖体小亚基(18S rRNA)编码基因的克隆与序列分析
黄勃, 方再光, 刘均玲, 周智, 王小兵
海南大学热带生物资源教育部重点实验室 海洋学院 海口570228
摘要:
采用分子生物学的方法,对南海不同海区的两个地理群体耳鲍(Haliotis asinina)18S rRNA基因全长进行了克隆和序列分析,并将耳鲍18S rRNA基因的序列与NCBI数据库中已收录鲍的18SrRNA基因进行了比较。结果发现,南海耳鲍核糖体18S rRNA基因与耳鲍H.asinina isolate H11核糖体18S rRNA基因序列的相同率高达98%;同一地理群体内耳鲍核糖体18S rRNA基因序列完全一致;不同地理群体间耳鲍核糖体18S rRNA基因在碱基组成上的相似率为99%,仅在某些位点处发生了碱基替换,即腺嘌呤(T)被鸟嘌呤(G)替换;同时,将这两个不同群体中耳鲍的18S rRNA基因与泰国耳鲍18S rRNA基因序列进行比较分析发现,它们之间也只是发生了碱基替换。
关键词:  地理群体  耳鲍  18S rRNA基因
DOI:
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基金项目:国家“863”资源环境技术领域项目,2004AA603130号;国家自然科学基金项目,40366001号;教育部科学技术研究重点项目资助,03146号;海南省外国专家局重点目资助,20031211号;海南大学海洋生物重点学科项目资助,200602号。
CLONING AND SEQUENCING OF GENES ENCODING HALIOTIS ASININA 18S rRNAs
HUANG Bo, FANG Zai-Guang, LIU Jun-Ling, ZHOU Zhi, WANG Xiao-Bing
Tropical Bio-resources of Education Department Key Laboratory, Ocean College, Hainan University, Haikou, 570228
Abstract:
Genetic differences of Haliotis asinina from 2 different geographic groups in the South China Sea were studied. One from the area near the Xisha Island and another near Hainan Island were analyzed by polymerase chain reaction (PCR) of 18S rRNA genes. The amplified 18S rRNA genes of individuals from the 2 groups were cloned and sequenced, and then aligned with other abalone 18S rRNA genes registered with NCBI GenBank. Results indicated that 18S rRNA genes of individuals from the same group had no differentiation, the similarity in the 18S rRNA genes neared 100%, whereas partial differentiation between the 2 groups was observed with the similarity up to 99%, and basal substitution took place at some sites. A neighbor-joining tree was constructed from sequence divergence of 18S rRNA genes, which all was built up by locating accurately those sequences according to species of abalone. An unweighted pair-group dendrogram method with arithmetic mean was constructed from divergence among the individuals from the 2 groups. The H. asinina in China and Thailand were genetically closer than other species of abalones, revealing a close relation in phylogeny among them.
Key words:  Geographic group, Haliotis asinina, 18S rRNA gene
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