Cen Xiaodong, Bi Qun, Zhu Shenggeng
College of Life Sciences, Peking University, Beijing 100871, People's Republic of China.
Appl Microbiol Biotechnol. 2006 Aug;71(5):767-72. doi: 10.1007/s00253-006-0334-5. Epub 2006 May 13.
Capacity and diversity are extremely important to the quality of various phage display libraries. In this work, lambda phage-based in vitro package was applied to construct a filamentous phage display antibody library so as to enlarge its capacity and introduce more sequence diversity in the final library. In vivo recombination via Cre recombinase/lox sites was also exploited to create V(H)/V(L) combination diversity based on multivalent package of lambda phage packaging extracts on phagemid DNA concatemers. The library constructed with 10 microg concatenated phagemid DNA and ten vials of lambda phage packaging extracts was calculated to contain 1.40 x 10(10) independent clones. Higher capacity can be easily achieved when more materials are consumed. This strategy is somewhat more efficient than prior methods.
库容和多样性对于各种噬菌体展示文库的质量极为重要。在本研究中，基于λ噬菌体的体外包装被用于构建丝状噬菌体展示抗体文库，以扩大其库容并在最终文库中引入更多的序列多样性。还利用通过Cre重组酶/lox位点的体内重组，基于λ噬菌体包装提取物对噬菌粒DNA串联体的多价包装来产生V(H)/V(L)组合多样性。用10μg串联的噬菌粒DNA和十管λ噬菌体包装提取物构建的文库经计算含有1.40×10(10)个独立克隆。消耗更多材料时可轻松实现更高的库容。该策略比先前的方法更有效。
