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The sensibility and specificity of the method were evaluated with the amplification and by sequencing randomized Rh-negative and Rh- positive controls.

序列特异性引物方法扩增RHD 基因特异的外显子及其侧翼序列，PCR产物直接序列分析测定核苷酸的变异。

The apxⅢA gene of Actinobacillus pleuropneumonie was amplified by PCR. The amplified DNA fragment 3 466bp was cloned into pMD18-T. After R.E. analysis and sequencing, the apxⅢA gene in pMD18-T was ligated into pBluescripⅡSK, the recombinant expression plasmid pET-28b/apxⅢA was constructed and analysed with R.E., the protein of apxⅢA gene expressed in E.coli BL21 was detected by Western blotting.

因胸膜肺炎放线杆菌的致病性主要是由毒素决定的，故参照猪胸膜肺炎放线杆血清2型菌株的序列(GenBankL1 2 1 4 5)设计了一对特异性引物，用PCR的方法扩增apxⅢA基因并得到了长 3 466bp的片段，然后将其克隆到pMD 1 8T中，经酶切鉴定和序列分析表明克隆是成功的；再将apxⅢA插入到原核表达载体pET 2 8b后，转化BL2 1 (DE3)，在IPTG诱导下获得高效表达，经Westernblotting检测证实表达产物有活性。

May be considered the equivalent of 5.9 rock in terms of relative technical ability required.

有很多85至90度冰壁艰苦的绳距，差不多是攀岩5.9所需要的技巧及能力。

Yes. Please have a seat, and we'll call you.

是的。请坐，我会叫您的。