西施舌16S rRNA基因片段PCR扩增及其核苷酸序列分析

利用酚/氯仿抽提法提取了西施舌闭壳肌总DNA,以总DNA为模板,利用两条16S rRNA基因特异引物,进行PCR扩增,扩增产物测序后进行序列分析。结果从西施舌的闭壳肌提取得到总DNA,获得一个长度约300 bp的PCR扩增产物;测序结果显示此片段为294 bp;核苷酸序列经WU-blast2分析,与Spisula subtruncata(AJ548774)和Corbicula sp(AY097259)的16S rRNA的同源性均为74%。经DNAStar分析,此核苷酸序列A,G,T,C含量分别为35.74%,25.51%,26.19%和13.59%,A T(61.90%)含量明显高于G C(38.10%)含量。

16S rRNA Gene Segment Amplification and Nucleotide Acid Sequence Analysis of Coelomactra antiquata

The total genomic DNA of Coelomactra antiquata(from the adductor muscle) was isolated using phenol/chloroform method,and it was used for the template of PCR.Two special primers for 16S rRNA gene fragment were designed and the PCR was carried out.The PCR product was sequenced and the nucleotide acids of 16S rRNA were analyzed using the DNAStar software package,and the identity was analyzed by Blastn(GenBank) and WU-blast2(EMBL).As a result,we obtained the total DNA from the adductor muscle,about 300 base-pair nucleotide acid sequences were acquired from the PCR,the PCR products was then sequenced and the 16S rRNA fragment of 294 bp was obtained.The result of WU-Blast demonstrated that the sequence identity is all 74% between Coelamactra antiquata and Spisula subtruncata(AJ548774),Coelomactra antiquata and Corbicula sp(AY097259).The contents of base A,G,T,C of the 16S rRNA gene fragment were 35.74%,25.51%,26.19% and 13.59% respectively,and the content of A T(61.90%) was obviously higher than that of G C(38.10%) in the nucleotide sequence of Coelomactra antiquata.