| 1. | Prior to the vector construction , the availability of gene replacement was extensively tested in streptomyces . sp . fr - 008
在载体构建之前，我们先在链霉菌fr - 008中验证了基因置换方法的可行性。 |
| 2. | Such deficiencies cause lysosomal storage disorders , such as gaucher ' s disease , and replacement of the gene is an effective treatment
这样的缺陷引起溶酶体堆积病（例如，高歇病） ，基因置换是一种有效的疗法。 |
| 3. | 2kb or 7 . 2kb regions abolished the productivity of antibiotic fr - 008 . the expected strain after gene replacement in the region of 5 . 2kb have not been obtained for unknown reason
尽管由于未知的原因尚未获得5 . 2kb区域的基因置换菌株，但是以上的试验证明基因置换过程是可以在链霉菌fr - 008中进行的。 |
| 4. | Phz621 - phz622 , the new gene replacement vector system , had been constructed through the insertion of the 1 . 0kb and 1 . 4kb flanking sequences of hau3r gene from wild - type s . lividans in the same natural orientation
预期在携带有大插入片段的基因置换yac分子进入野生型变铅青链霉菌后， yac分子将通过1 . 4kb或1 |
| 5. | Lividans mutant strains la1 and la2 generated by gene replacement in dndb and dndc were obtained respectively . both of these two mutants lost dnd phenotype because of disruption of a respective dnd gene
首先利用基因置换实验获得分别在dndb 、 dndc基因内部发生基因中断的突变菌株la1利la2 ，并通过southern杂交进行了验证。 |
| 6. | Secondly another vector phz1117 which for replacement of sc ( j11 . 17 / scd78 . 01 , was constructed . phz1117 was transformed into wild type strain m145 and a gene replacement strain named hxy3 was obtained
其次，构建了另一个用于置换scq11 . 17 scd78 . 01的置换载体phz1117 ，得到基因置换菌株hxy3并通过southern杂交验证，与野生型m145相比，没有观察到hxy3有明显的表型变化。 |
| 7. | In order to facilitate the study of biological function of add gene cluster , e . coli - s . avermitilis intragenus conjugation system was established . in addition , phz2114 for the replacement of the entire add gene cluster and phz2130 for disruption of adda were constructed
建立了除虫链霉菌的接合转移系统，并构建了用于置换全部基因簇的基因置换质粒phz2114和adda的基因中断质粒phz2130 ，为研究除虫链霉菌add基因簇的生物学功能奠定了基础。 |
| 8. | Pfge analysis of these 21 blocked strains revealed a common chromosomal deletion in the 300kb asel fragment which might be responsible for antibiotic 5102 - iii biosynthesis . so the 300kb fragment was recovered and used as probe to hybridize with 10 - 22 genomic library . cosmids in this region were aligned and suitable fragments in this region were selected and used further for the construction of gene replacement plasmids
以此片段为探针钓出了位于该区域的文库克隆，并利用指纹印迹和杂交技术将这些文库克隆排列起来，进而以位于该区域的不同位置的片段做臂构建了用于转化和接合转移用的基因置换质粒，并试图通过基因置换将该区域置换下来，但尚未得到最终结果。 |
| 9. | The complicated life cycle of streptomyccs is one of the most important reason that attracts researchers on its genetics . a great deal of valuable information on the morphological , physiological differentiation of streptomyccs coclicolor will emerge after genome sequencing is completed . based on the s . coelicolor dna sequences that have been finished and focusing on three ftsk homologous , which may be closely related with cell division , this study tried to find out the functions of three ftsk homologues genes by gene replacement or disruption
链霉菌复杂的生活史是研究其遗传学的一个重要原因，天蓝色链霉菌全基因组测序将会为其形态及生理分化发育的研究提供大量有用的信息，本研究基于天蓝色链霉菌已测出的98序列，对一组可能与细胞分裂有关的基因? ftdk同源基因? ?进行基因置换或中断并对其功能进行初步研究。 |
| 10. | To facilitate the detection of gene replacement , apr gene was placed in the middle of the two inserts . orit from e . coli plasmid rk2 was also incoporated into the vector , therefore , pij903 derivatives can be mobilized from e . coli into streptomyces sp . fr - 008 by either bi - parental conjugation or by conjugation between streptomyces
为了在克隆完整基因簇时避免这两方面的影响并能随机地获得dna大片段，我们将第二代yac载体pjs97 - pjs98改造成为适合于在白林泉链霉菌基因组中dna大片段的基因置换与克隆链霉菌中分析被克隆大片段功能的基因置换载体。 |
