ABSTRACT
In this paper, Liuwei Dihuang pill was used to study the identification of Chinese patent medicine by fluorescence sequencing typing technology. The DNA of Paeonia suffruticosa was used as template to amplify by five pair of FAM fluorescence labeling primers. Then, the amplified products were sequenced. The sequencing results were analyzed by GeneMarker V1.80 to screen the best fluorescence labeling primers. As a result, psbA-trnH fluorescence labeling primer was used to identify the raw materials of Liuwei Dihuang pill. The results showed that three kinds of raw plant medicinal materials in Liuwei Dihuang pill were able to be correctly identified by psbA-trnH fluorescence labeling primer. The fluorescence sequencing typing technology can stably and accurately distinguish raw medicinal materials in Chinese patent medicine.
Subject(s)
DNA Primers , Chemistry , Genetics , DNA, Plant , Chemistry , Genetics , Drugs, Chinese Herbal , Chemistry , Reference Standards , Fluorescent Dyes , Chemistry , Plants, Medicinal , Chemistry , Genetics , Polymerase Chain Reaction , Methods , Quality Control , Staining and LabelingABSTRACT
<p><b>OBJECTIVE</b>To explore the new method of discriminating Gelsemium elegans from Lonicera japonica and its close species by using specific PCR amplification.</p><p><b>METHOD</b>Thirteen samples of the different G. elegans materials and 58 samples of L. japonica, L. macranthoides and L. dasystyla were collected. The total DNA of the samples were extracted, and the DNA of G. elegans, L. japonica and L. macranthoides water extracts were extracted. PsbA-rnnH sequence from G. elegans was amplified by PCR and sequenced unidirectionally, ClustulW was used to align psbA-trnH sequences of the G. elegans and L. japonica and its close species from GenBank database.</p><p><b>RESULT</b>All samples were amplified by PCR with specific primer, DNA from G. elegans would be amplified 97 bp whereas PCR products from all of Lonicera samples had not bands.</p><p><b>CONCLUSION</b>Specific PCR amplification can be used to identify G. elegans from L. japonica and its close species successfully and is an efficient molecular marker for authentication of G. elegans and L. japonica and its close species.</p>
Subject(s)
DNA, Plant , Genetics , Drugs, Chinese Herbal , Gelsemium , Chemistry , Genetics , Lonicera , Chemistry , Genetics , Phylogeny , Plant Extracts , Chemistry , Genetics , Polymerase Chain Reaction , Water , ChemistryABSTRACT
To explore the new method of discriminating Astragali Radix and Hedysari Radix by using PCR amplification of specific alleles, 30 samples of the different Astragali Radix materials and 28 samples of Hedysari Radix were collected. The total DNA of all samples were extracted, trnL-trnF sequence from Astragali Radix and Hedysari Radix was amplified by PCR and sequenced unidirectionally. These sequences were aligned by using Clustul W. Primer was designed and the PCR reaction systems including annealing temperature, dNTP, etc were optimized. All samples were amplified by PCR with specific primer, DNA from Astragali Radix would be amplified 136 bp, whereas PCR products from all of Hedysari Radix were 323 bp. This method can detect 10% of intentional Hedysari Radix DNA into Astragali Radix. PCR amplification of alleles can be used to identify Astragali Radix and Hedysari Radix successfully and is an efficient molecular marker for authentication of Astragali Radix and Hedysari Radix.
Subject(s)
Alleles , Astragalus Plant , Classification , Genetics , DNA Barcoding, Taxonomic , DNA, Plant , Genetics , Polymerase Chain ReactionABSTRACT
Lian Qiao Bai Du Wan was used to study the identification of Chinese patent medicine by molecular marker technique. DNA was extracted through modified CTAB method. The psbA-trnH and rbcL sequences were gradient amplified, and PCR products were ligated with the pEASY-T5 vector and then transformed into Trans1-T1 cells, respectively. Clones were selected randomly and sequenced. All sequences were analyzed by BlastN and the neighbor-joining (NJ) phylogenetic tree was constructed by MEGA 4.0. The results showed that nine kinds of medicinal materials can be identified by psbA-trnH sequences, and six kinds of medicinal materials by rbcL sequences from Lian Qiao Bai Du Wan. Molecular marker technique can stably and accurately distinguish multi-origin medicinal materials in Chinese patent medicine.
Subject(s)
Base Sequence , Chloroplasts , Genetics , Cluster Analysis , DNA Barcoding, Taxonomic , DNA, Chloroplast , Genetics , DNA, Intergenic , Genetics , DNA, Plant , Genetics , Drugs, Chinese Herbal , Chemistry , Forsythia , Chemistry , Genetics , Phylogeny , Plants, Medicinal , Chemistry , Genetics , Polymerase Chain Reaction , Ribulose-Bisphosphate Carboxylase , Genetics , Sequence Analysis, DNA , Species SpecificityABSTRACT
<p><b>OBJECTIVE</b>To explore a new method for identification Astragali Radix from its adulterants by using ITS sequence.</p><p><b>METHOD</b>Thirteen samples of the different Astragali Radix materials and 6 samples of the adulterants of the roots of Hedysarum polybotrys, Medicago sativa and Althaea rosea were collected. ITS sequence was amplified by PCR and sequenced unidirectionally. The interspecific K-2-P distances of Astragali Radix and its adulterants were calculated, and NJ tree and UPGMA tree were constructed by MEGA 4.</p><p><b>RESULT</b>ITS sequences were obtained from 19 samples respectively, there were Astragali Radix 646-650 bp, H. polybotrys 664 bp, Medicago sativa 659 bp, Althaea rosea 728 bp, which were registered in the GenBank. Phylogeny trees reconstruction using NJ and UPGMA analysis based on ITS nucleotide sequences can effectively distinguish Astragali Radix from adulterants.</p><p><b>CONCLUSION</b>ITS sequence can be used to identify Astragali Radix from its adulterants successfully and is an efficient molecular marker for authentication of Astragali Radix and its adulterants.</p>
Subject(s)
Althaea , Classification , Genetics , Astragalus propinquus , Classification , Genetics , DNA, Plant , Chemistry , Genetics , DNA, Ribosomal , Chemistry , Genetics , DNA, Ribosomal Spacer , Genetics , Fabaceae , Classification , Genetics , Medicago sativa , Classification , Genetics , Molecular Sequence Data , Phylogeny , Plant Roots , Genetics , RNA, Ribosomal , Genetics , Genetics , Sequence Analysis, DNA , Species SpecificityABSTRACT
<p><b>OBJECTIVE</b>To investigate the polymorphism distribution of vitamin D receptor (VDR) gene Fok I in Mongolian population of China.</p><p><b>METHODS</b>Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to analyze three genotypes FF, Ff and ff in the start codon of VDR gene (Fok I) in unrelated normal healthy Mongolian individuals of China.</p><p><b>RESULTS</b>In the population, we obtained the allelic frequencies of 57% and 43% for (F) and (f) allele and the percentage of genotypes FF, Ff and ff to be 31%, 52%, and 17% respectively.</p><p><b>CONCLUSION</b>The polymorphism frequency and distribution of this VDR gene Fok I in Mongolian population of China exhibit its own characteristics.</p>