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Molecular analysis of the 16S-23S rDNA internal spacer region (ISR) and truncated tRNA(Ala) gene segments in Campylobacter lari

机译:弯曲杆菌中16S-23S rDNA内部间隔区(ISR)和tRNA(Ala)基因截短的分子分析

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摘要

Following PCR amplification and sequencing, nucleotide sequence alignment analyses demonstrated the presence of two kinds of 16S-23S rDNA internal spacer regions (ISRs), namely, long length ISRs of 837-844 base pair (bp) [n = six for urease-negative (UN) isolates, UN JCM2530(T), RM2100, 176, 293, 299 and 448] and short length ISRs of 679-725 bp [n = six for UN n = 14 for urease-positive thermophilic (UPTC) isolates]. The analyses also indicated that the short length ISRs mainly lacked the 156 bp sequence from the nucleotide positions 122-277 bp in long length ISRs for UN JCM2530(T). The 156 bp sequences shared 94.9-96.8 % sequence similarity among six isolates. Surprisingly, atypical tRNA(Ala) gene segment (5' end 35 bp), which was extremely truncated, occurred within the 156 bp sequences in the long length ISRs, as an unexpected tRNA(Ala) pseudogene. An order of the intercistronic tRNA genes within the short nucleotide spacer of 5'-16S rDNA-tRNA(Ala)-tRNA(Ile)-23S rDNA-3' occurred in all the isolates examined.
机译:经过PCR扩增和测序后,核苷酸序列比对分析表明存在两种16S-23S rDNA内部间隔区(ISR),即837-844个碱基对的长ISR(bp)[对于尿素酶阴性,n = 6 (UN)分离株,UN JCM2530(T),RM2100、176、293、299和448]和679-725 bp的短长度ISR [对于尿素酶阳性嗜热(UPTC)分离株,n = 6,UN n = 14]。分析还表明,UN JCM2530(T)的长ISR中,短ISR的核苷酸位置122-277 bp中主要缺少156 bp序列。六个分离株之间的156 bp序列共有94.9-96.8%的序列相似性。出人意料的是,非典型的tRNA(Ala)基因片段(5'端35 bp)被极大地截短,出现在长ISR的156 bp序列内,是一个意外的tRNA(Ala)假基因。在所有检测到的分离物中,在5'-16S rDNA-tRNA(Ala)-tRNA(Ile)-23S rDNA-3'的短核苷酸间隔区中存在顺反子间tRNA基因的顺序。

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