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COI 염기서열 기반 백강잠 신속 감별용 SCAR marker 개발 - 백강잠 유전자 감별 -

Development SCAR marker for the rapid authenticatonof Batryticatus Bombyx based on COI Sequences

대한본초학회지 2019년 34권 5호 p.13 ~ 20
김욱진 ( Kim Wook-Jin ) - 한국한의학연구원 한약자원연구센터

양선규 ( Yang Sun-Gyu ) - 한국한의학연구원 한약자원연구센터
노푸름 ( Noh Pu-Reum ) - 한국한의학연구원 한약자원연구센터
박인규 ( Park In-Kyu ) - 한국한의학연구원 한약자원연구센터
최고야 ( Choi Go-Ya ) - 한국한의학연구원 한약자원연구센터
송준호 ( Song Jun-Ho ) - 한국한의학연구원 한약자원연구센터
문병철 ( Moon Byeong-Cheol ) - 한국한의학연구원 한약자원연구센터

Abstract


Objectives : To ensure the safety, quality and pharmacological efficacy of Batryticatus Bombyx, it is important to discriminate with adulterants. In Korean Herbal Pharmacopoeias (KHP), the authentic species of Batryticatus Bombyx is defined only Bombyx mori. Therefore, the aim of this study is establishment of PCR assay method using the sequence characterized amplified region (SCAR) marker based on COI DNA barcode for discriminating six species related to Batryticatus Bombyx.

Methods : Seventeen samples of six species (Bombyx mori, Bombyx mandarina, Rhodinia fugax, Oberthueria caeca, Actias artemis, and Caligula japponica) were collected from different habitate and nucleotide sequences of cytochrome c oxidase subunit I(COI) barcode regions were analyzed by Sanger sequencing methods. To develop SCAR-based PCR assay method, we designed species-specific primers based on COI sequence variabilities and verified those specificities using 17 samples of six species as well as commercial herbal medicines.

Results : In comparative multiple analysis of COI sequences, six species were distinguished by species-specific nucleotides at the species level. To develop rapid and reliable PCR assay method for genetic authentication of Batryticatus Bombyx, therefore, we designed species-specific SCAR primers based on these nucleotide sequences and confirmed those specificities. Using these SCAR primers, We also established simple conventional PCR assay method using these SCAR primers at the species level.

Conclusions : The comparative analysis of COI sequences and SCAR-based PCR assay methods represented equal results for distinguishing authentic Batryticatus Bombyx and adulterations at the species level. Therefore, our results are expected protecting adulteration of herbal medicine Batryticatus Bombyx.

키워드

Bombyx mori; Batryticatus Bombyx; COI; DNA barcoding; SCAR marker
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