Sequence analysis of chloroplast chlB gene of medicinal Ephedra species and its application to authentication of Ephedra Herb.
Chloroplast chlB gene encoding subunit B of light-independent protochlorophyllide reductase was amplified from herbarium and crude drug specimens of Ephedra sinica, E. intermedia, E. equisetina, and E. przewalskii. Sequence comparison of the chlB gene indicated that all the E. sinica specimens have the same sequence type (Type S) distinctive from other species, while there are two sequence types (Type E1 and Type E2) in E. equisetina. E. intermedia and E. prezewalskii revealed an identical sequence type (Type IP). E. sinica was also identified by digesting the chlB fragment with Bcl I. A novel method for DNA authentication of Ephedra Herb based on the sequences of the chloroplast chlB gene and internal transcribed spacer of nuclear rRNA genes was developed and successfully applied for identification of the crude drugs obtained in the Chinese market.
Guo
Yahong
Y
Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan.
Tsuruga
Ayako
A
Yamaguchi
Shigeharu
S
Oba
Koji
K
Iwai
Kasumi
K
Sekita
Setsuko
S
Mizukami
Hajime
H
D001483:Base Sequence; D002736:Chloroplasts / Q000201:enzymology / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D004365:Drugs, Chinese Herbal / Q000737:chemistry* / Q000592:standards; D029789:Ephedra / Q000737:chemistry / Q000145:classification* / Q000201:enzymology / Q000235:genetics*; D017343:Genes, Plant; D020835:Medicine, Kampo; D008969:Molecular Sequence Data; D044925:Oxidoreductases Acting on CH-CH Group Donors / Q000737:chemistry* / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA
D018744:DNA, Plant; D004365:Drugs, Chinese Herbal; D044925:Oxidoreductases Acting on CH-CH Group Donors; C023886:protochlorophyllide reductase