AIM To analyze sequences of the nuclear ribosomal RNA small subunit (18S rRNA) gene and the chloroplast mat K gene of crude drug Patchouli ( Pogostemon cablin ) in order to provide molecular evidence for identificatio...AIM To analyze sequences of the nuclear ribosomal RNA small subunit (18S rRNA) gene and the chloroplast mat K gene of crude drug Patchouli ( Pogostemon cablin ) in order to provide molecular evidence for identification of Patchouli drug. METHODS To sequence the entire 18S rRNA gene and partial mat K gene of Patchouli from Guangzhou and its substitute Wrinkled Gianthyssop ( Agastache rugosa ) from S ichuan using PCR direct sequencing and to detect the homology of two gene sequen ces between these two crude drugs. RESULTS The complete 18S rRNA gene sequence is 1 805 bp in length for Patchouli from Guangzhou whereas 1 794 bp for Wrinkled Gianthyssop from Sichuan. The 3′ end sequence of mat K gene is 521 bp (747～ 1 268 nt from upstream of ma t K gene) for these two crude drugs. Based on multiple sequence alignment, it i s found that there are 18 variable sites and 11 aligned gap sites in 18S rRNA se quence, 49 variable sites in 3′ mat K sequence between these two crude drug s. The homology is 98 4% for 18S rRNA and 90 6% for 3′ mat K between two crude drugs, respectively. CONCLUSION DNA sequencing can provide an accurate and reliable tool in the crude drug ident ification of Patchouli and its substitute Wrinkled Gianthyssop.展开更多
自新疆伊宁地区黄牛体表采集的盾糙璃眼蜱饥饿或半饱血成虫,带回实验室感染除脾牛。经血涂片检查发现,于感染后第21天,病牛血液中出现一种以圆环形与卵圆形为主的泰勒虫。经形态学观察和18 S rRNA基因测序和进化关系分析证明,它们为环...自新疆伊宁地区黄牛体表采集的盾糙璃眼蜱饥饿或半饱血成虫,带回实验室感染除脾牛。经血涂片检查发现,于感染后第21天,病牛血液中出现一种以圆环形与卵圆形为主的泰勒虫。经形态学观察和18 S rRNA基因测序和进化关系分析证明,它们为环形泰勒虫。将此饱血成蜱经实验室饲喂条件下,孵化出的幼虫分别饲喂于牛与家兔两种不同的动物,结果在饱血若虫阶段所有的蜱均从两种动物体表脱落,不再叮咬,饥饿成蜱又更换另一宿主,证明其是二宿主蜱。展开更多
参照GenBank收录的Isospora suis 18SrRNA基因全序列(U97523),利用引物分析软件Oligo 6.57和Primer Premier 5.0设计了一对引物(上游引物:5'-tcctgcgagtactcatatgc-3';下游引物:5'-gttcagc-tacgcataccttg-3'),首次扩增...参照GenBank收录的Isospora suis 18SrRNA基因全序列(U97523),利用引物分析软件Oligo 6.57和Primer Premier 5.0设计了一对引物(上游引物:5'-tcctgcgagtactcatatgc-3';下游引物:5'-gttcagc-tacgcataccttg-3'),首次扩增出猪等孢球虫分离株的18SrRNA基因序列,结合GenBank上登录的相关原虫序列,用DNAstar4.0软件比较其同源性,经Clustalx1.81序列比对,Paup4.0、Treeview3.0、Phylip种系发育关系软件分析后,证实该分离株为猪等孢球虫,并且河南不同地区之间的猪等孢球虫没有明显遗传差异。展开更多
文摘AIM To analyze sequences of the nuclear ribosomal RNA small subunit (18S rRNA) gene and the chloroplast mat K gene of crude drug Patchouli ( Pogostemon cablin ) in order to provide molecular evidence for identification of Patchouli drug. METHODS To sequence the entire 18S rRNA gene and partial mat K gene of Patchouli from Guangzhou and its substitute Wrinkled Gianthyssop ( Agastache rugosa ) from S ichuan using PCR direct sequencing and to detect the homology of two gene sequen ces between these two crude drugs. RESULTS The complete 18S rRNA gene sequence is 1 805 bp in length for Patchouli from Guangzhou whereas 1 794 bp for Wrinkled Gianthyssop from Sichuan. The 3′ end sequence of mat K gene is 521 bp (747～ 1 268 nt from upstream of ma t K gene) for these two crude drugs. Based on multiple sequence alignment, it i s found that there are 18 variable sites and 11 aligned gap sites in 18S rRNA se quence, 49 variable sites in 3′ mat K sequence between these two crude drug s. The homology is 98 4% for 18S rRNA and 90 6% for 3′ mat K between two crude drugs, respectively. CONCLUSION DNA sequencing can provide an accurate and reliable tool in the crude drug ident ification of Patchouli and its substitute Wrinkled Gianthyssop.
文摘自新疆伊宁地区黄牛体表采集的盾糙璃眼蜱饥饿或半饱血成虫,带回实验室感染除脾牛。经血涂片检查发现,于感染后第21天,病牛血液中出现一种以圆环形与卵圆形为主的泰勒虫。经形态学观察和18 S rRNA基因测序和进化关系分析证明,它们为环形泰勒虫。将此饱血成蜱经实验室饲喂条件下,孵化出的幼虫分别饲喂于牛与家兔两种不同的动物,结果在饱血若虫阶段所有的蜱均从两种动物体表脱落,不再叮咬,饥饿成蜱又更换另一宿主,证明其是二宿主蜱。
文摘参照GenBank收录的Isospora suis 18SrRNA基因全序列(U97523),利用引物分析软件Oligo 6.57和Primer Premier 5.0设计了一对引物(上游引物:5'-tcctgcgagtactcatatgc-3';下游引物:5'-gttcagc-tacgcataccttg-3'),首次扩增出猪等孢球虫分离株的18SrRNA基因序列,结合GenBank上登录的相关原虫序列,用DNAstar4.0软件比较其同源性,经Clustalx1.81序列比对,Paup4.0、Treeview3.0、Phylip种系发育关系软件分析后,证实该分离株为猪等孢球虫,并且河南不同地区之间的猪等孢球虫没有明显遗传差异。
