Mutation characteristics of rpoB gene in rifampicin-resistantBrucella strains
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目的 分析布鲁氏菌利福平耐药株rpoB基因的突变特征。 方法 选取分离自新疆维吾尔自治区的4株布鲁氏菌利福平耐药株（JSY-26、G-9、WSY-13、AW-3株）DNA，采用PCR扩增利福平rpoB基因，并对核苷酸序列进行测序。以布鲁氏菌利福平耐药标准株（RB51株）和利福平敏感株（ALT-8株）的rpoB基因序列为参照，应用Mega 7.0软件比对分析4株布鲁氏菌利福平耐药株rpoB基因利福平耐药决定区（rifampicin resistance determination region，RRDR）内外的碱基突变位点及类型。 结果 经序列比对分析，JSY-26和WSY-13株均在rpoB基因RRDR 1 576 bp处发生单碱基点突变，碱基由鸟嘌呤（guanine，G）突变为腺嘌呤（adenine，A）；G-9株在rpoB基因RRDR 1 606 bp处发生单碱基点突变，碱基由胞嘧啶（cytosine，C）突变为A。AW-3株在rpoB基因RRDR外2 536、2 537、2 626、2 636、2 654 bp处出现3种类型共5处突变，分别为插入突变3处[胸腺嘧啶（thymine，T）插入1次、C插入2次]，缺失突变1处（C缺失），单碱基点突变1处（由G突变为C）。 结论 布鲁氏菌利福平耐药株rpoB基因RRDR突变以单碱基点突变为主，RRDR外则出现多位点插入和缺失突变。 Objective To analyze the mutation characteristics of rpoB gene in rifampicin-resistant Brucella strains. Methods DNA of 4 rifampicin-resistant Brucella strains (JSY-26, G-9, WSY-13 and AW-3) isolated from Xinjiang Uygur Autonomous Region was selected, rifampicin rpoB gene was amplified by PCR and its nucleotide sequence was sequenced. The rpoB gene sequences of rifampicin-resistant Brucella standard strain (RB51) and sensitive strain (ALT-8) were used as reference, the mutation sites and types of the rpoB gene inside and outside the rifampicin resistance determination region (RRDR) of the 4 rifampicin-resistant Brucella strains were analyzed by Mega 7.0 software. Results Through sequence alignment, both JSY-26 and WSY-13 strains underwent a single base point mutation at the RRDR 1 576 bp of the rpoB gene, with the base changing from guanine (G) to adenine (A). The G-9 strain underwent a single base point mutation at the RRDR 1 606 bp of the rpoB gene, with the base changing from cytosine (C) to A. The AW-3 strain showed 5 mutations of 3 types outside rpoB gene RRDR at 2 536, 2 537, 2 626, 2 636 and 2 654 bp, namely 3 insertion mutations [thymine (T) insertion once and C insertion twice], 1 deletion mutation (C deletion), and 1 single base point mutation (from G to C mutation). Conclusion The RRDR mutations in the rpoB gene of the rifampicin-resistant Brucella strains are mainly characterized by single base point mutations, while multiple insertion and deletion mutations occur outside the RRDR.
Objective To analyze the mutation characteristics of rpoB gene in rifampicin-resistant Brucella strains. Methods DNA of 4 rifampicin-resistant Brucella strains (JSY-26, G-9, WSY-13 and AW-3) isolated from Xinjiang Uygur Autonomous Region was selected, rifampicin rpoB gene was amplified by PCR and its nucleotide sequence was sequenced. The rpoB gene sequences of rifampicin-resistant Brucella standard strain (RB51) and sensitive strain (ALT-8) were used as reference, the mutation sites and types of the rpoB gene inside and outside the rifampicin resistance determination region (RRDR) of the 4 rifampicin-resistant Brucella strains were analyzed by Mega 7.0 software. Results Through sequence alignment, both JSY-26 and WSY-13 strains underwent a single base point mutation at the RRDR 1 576 bp of the rpoB gene, with the base changing from guanine (G) to adenine (A). The G-9 strain underwent a single base point mutation at the RRDR 1 606 bp of the rpoB gene, with the base changing from cytosine (C) to A. The AW-3 strain showed 5 mutations of 3 types outside rpoB gene RRDR at 2 536, 2 537, 2 626, 2 636 and 2 654 bp, namely 3 insertion mutations [thymine (T) insertion once and C insertion twice], 1 deletion mutation (C deletion), and 1 single base point mutation (from G to C mutation). Conclusion The RRDR mutations in the rpoB gene of the rifampicin-resistant Brucella strains are mainly characterized by single base point mutations, while multiple insertion and deletion mutations occur outside the RRDR.
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