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
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
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
基金Supported by Key Project of Knowledge Innovation Project of Chinese Academy of Sciences(KGCX2-YW-374-3)Scientific and Technological Project of Shandong Province(2008GG20007002)~~
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。
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。
