异序的

Pneumoniae FH strain was cloned and the sequence was analysed by M13 DNA sequencing method. Comparing the PCR product sequcence with MP M-129 strain P1 gene, we found that there are 4 bases different. This may result from the different MP DNA templates. The maximum homology is 98.8%. The result confirmed the fidelity and specificity of the amplified target DNA segment by PCP, and suggested that two categories of MP P1 gene still exist a few differences even in the conservation region. The cloning MP DNA segment was labelled by random hexanucleotide priming, after hybridization, the probe detection was completed using an anti-digoxigenin antibody alkaline phosphatase conjugate, and the substrates 5-bromo-4-chloro-3-indolyl phosphate and nitro blue tetrazolium. This hybridization system is much superior to the radioactive probe hybridization, because it is safe, easy to handle and has no limitation of decay time. The time required for colormetic detection is also much less than the corresponding autoradiographic exposure time needed to achieve similar detection limits with 32P-labelled probes. The Dig-probes could be used repeatedly, and this made them not only much convenient to use, but also lower the cost, and worthwhile to be used popularly.

将PCR产物进行重组，并将阳性重组质粒，应用M13测序系统对产物进行DNA序列分析，并与MPM-129株P1基因核苷酸进行同源性比较，发现有4个位置的碱基发生了变化，其同源性为98.8％，证实了PCR所扩增DNA片段的准确性和特异性，同时也证实了不同MP组型的P1基因即使在保守区也存在着一定的差异，将克隆的目的DNA片段用异羟基洋地黄毒苷配基用随机引物法标记制备MP DNA探针，杂交后用碱性磷酸酶标记的抗Dig多克隆抗体与杂交体反应，再用BCIP和NBT呈色，制备MP DNA探针，鉴定所扩增片段的特异性，与同位素探针比较，Dig探针不受半衰期限制，可反复使用，而且价格低廉，值得推广使用。

As she looked at Warrington's manly face, and dark, melancholy eyes, she had settled in her mind that he must have been the victim of an unhappy attachment.

每逢看到沃林顿那刚毅的脸，那乌黑、忧郁的眼睛，她便会相信，他一定作过不幸的爱情的受害者。

Maybe they'll disappear into a pothole.

也许他们将在壶穴里消失