| 粟绒焦淑静黄现恩张青史全良△.苏铁珊瑚根共生念珠藻ITS序列多拷贝研究*[J].,2014,14(15):2851-2857 |
| 苏铁珊瑚根共生念珠藻ITS序列多拷贝研究* |
| The Study on Multicopy of ITS Sequences of Nostoc in Cycads Revolute* |
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| DOI： |
| 中文关键词: 共生念珠藻 ITS序列 假阳性 协同进化 |
| 英文关键词: Symbiotic Nostoc ITS sequence False positive Concerted evolution |
| 基金项目:江苏省昆山科技局资助项目（N3134930） |
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| 中文摘要: |
| 摘要目的：研究苏铁珊瑚根共生念珠藻16SrDNA-23SrDNA 基因间隔序列（ITS）拷贝数。方法：提取苏铁珊瑚根共生念珠藻全
DNA，PCR 扩增其16SrDNA-23SrDNA基因间隔序列（ITS）克隆并测序；采用Cluster X 1.83对所得序列进行比对，用DNAman 软
件对比对结果进行人工校正；采用割胶回收电泳检测、菌液PCR及混合模板PCR 扩增三种方法验证ITS-L2。结果：ITS-S(420 bp)
不含任何氨基酸编码序列，ITS-L1(676 bp)包含一个异亮氨酸和一个丙氨酸编码序列；ITS-L1 和ITS-L2 在琼脂糖凝胶电泳检测
时长度分别为676 bp 和1000 bp，而谱带经克隆测序后结果表明ITS-L1 和ITS-L2 碱基序列和长度几乎相同，仅有两个碱基发生
变异。结论：ITS-L2 为假阳性，可能是由ITS-S 和ITS-L1 形成的异源双链；苏铁珊瑚根共生念珠藻含有两种类型的ITS拷贝，一
种是含有tRNAIle 和tRNAAla 编码序列的ITS，另一种是不含tRNAIle 和tRNAAla 编码序列的ITS。 |
| 英文摘要: |
| ABSTRACT Objective:To investigate the multicopy number of ITS sequences of Nostoc in . Methods:The total
DNA of Nostoc in was extracted. 16S rDNA-23SrDNA internal transcribed spacer (ITS) was amplified from ,
which is symbiont in cycads revolute, then was TA-cloned and sequenced. The sequences were blasted by Cluster X 1.83, and DNA man
was used to calibrate the results; Electrophoresis detection of PCR amplification with bacteria solution or mixture template was used to
check the results. Results:The PCR amplification had three products [420 bp (ITS-S),676 bp (ITS-L1),1000 bp (ITS-L2)]. And the
sequences indicated that none of the tRNA coding region was included in the ITS-S (420 bp), while ITS-L1 (676 bp) included both
tRNAIle and tRNAAla coding regions. ITS-L1 and ITS-L2 were at 676 bp and 1000 bp. But the sequences results showed they were
almost of the same length and base sequences, and only two bases were different. Conclusion:It was inferred that ITS-L2 was false
positive, and it maybe heteroduplexes composed of ITS-S and ITS-L1. There are two types of multicopy of ITS sequences of Nostoc in
cycads revolute. One includes both tRNAIle and tRNAAla coding region, and the other includes none. |
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