Acta Scientiarum Naturalium Universitatis Pekinensis

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Molecular Cloning and Sequencing the 18S rDNA from Specialized Dinosaur Egg Fossil Found in Xixia Henan, China

AN Chengcai, LI Yi, ZHU Yuxian, SHEN Xing, ZHANG Yun,YOU Lingtao, LIANG Xiaowen, LI Xiaohua, WU Si,WU Peng, GU Hongya, ZHOU Zengquan, CHEN Zhanglian   

  1. College of Life Sciences, Peking University, Beijing 100871
  • Received:1995-02-05 Online:1995-03-20 Published:1995-03-20

中国河南西峡恐龙蛋化石中18SrDNA部分片段的克隆及序列分析

安成才,李毅,朱玉贤,沈兴,张昀,由凌涛,梁晓文,李小华,吴思,伍鹏,顾红雅,周曾铨,陈章良   

  1. 北京大学生命科学学院,北京,100871

Abstract: We have extracted DNA from the inclusion of a dinosaur egg fossil from the Late Cretaceous in Xixia Henan, China. DNA fragments are isolated from the floccus in the cavity of the egg fossil, and confirmed by the method of random primer isotope labelling. Their size ranges from tens base pairs to one thousand base pairs, and most of them are less than four hundred base pairs. Polymerase chain reaction (PCR) is performed with the specific 18S rDNA primers and the extracted DNA as template, and DNA fragments about 200 base pairs are obtained. After ligation, transformation and screening, we get six positive clones. Sequence analysis and homology comparison reveal that these fragments have high homology with the 18S rDNA of Amphlbia, Reptile, Aves and Humanin order, but no significant homology with prokaryotes. The same sequenceshave not been found in the EMBL data bank.

Key words: dinosaur egg, Xixia, 18S rDNA, sequencing

摘要: 本文从河南省西峡晚白垩世的一枚保存方式特殊的恐龙蛋化石中提取DNA,经过32P同位素标记和电泳分析,发现在蛋内腔的絮状物样品中确有DNA片段存在。它们的大小约在1000bp至几十b[之间,大多数片段在400bp以下。用18SrDNA特异引物,以上述DNA为模板进行了PCR扩增,经电泳分析得到约200bp的PCR产物。经过连接、转化到大肠杆菌,最终筛选出6个阳性克隆。测序分析和同源性比较发现这6个克隆的序列与两栖类、爬行类、鸟类及人类等的18具有很高的同源性,但是与原核生物没有发现同源性,而且没有检索出与该基因完全相同的已知序列。

关键词: 恐龙蛋, 西峡, 18SrDNA, 序列测定

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