公开(公告)号：US20060147986A1

公开(公告)日：2006-07-06

申请号：US11369726

申请日：2006-03-07

申请人： Anna Hine ,  Jane Morgan ,  Albert Santos ,  David Palfrey

发明人： Anna Hine ,  Jane Morgan ,  Albert Santos ,  David Palfrey

IPC分类号： C40B40/08 ,  C40B40/10

CPC分类号： C12N15/1093 ,  C07K1/047 ,  C07K14/001 ,  C12N15/1034 ,  G01N33/6872

摘要： A set of libraries of proteins, which proteins are capable of specific binding interactions by virtue of amino acid residues at two or more determined positions including a first determined position and one or more other determined positions, which set of libraries consists of: a) 6 to 20 libraries in which each library has one or several but less than 20 amino acid residues at the said first determined position and is randomised at the said one or more other determined positions, the arrangement being such that interaction of the 6 to 20 libraries with a specific binding partner identifies an amino acid residue at the said first determined position that takes part in the specific binding interaction, and b) 6 to 20 libraries of corresponding design for each of the said one or more other determined positions. A set of libraries of genes which code for the proteins. A method of identifying a protein which interacts with a specific binding partner, which method comprises incubating the protein with each library of the set of libraries of proteins, observing specific binding interactions with certain libraries of the set, and using the observations to identify a protein which interacts with the specific binding partner. A method of making a library of randomised genes.

摘要翻译： 一组蛋白质库，该蛋白质通过两个或更多个确定位置处的氨基酸残基能够进行特异性结合相互作用，包括第一确定位置和一个或多个其他确定的位置，该组文库由以下组成：a）6 其中每个文库在所述第一确定位置具有一个或多个但少于20个氨基酸残基的20个文库，并且在所述一个或多个其他确定的位置被随机化，所述方案使得6至20个文库与 特异性结合配偶体识别参与特异性结合相互作用的所述第一确定位置处的氨基酸残基，以及b）对于所述一个或多个其它确定位置中的每一个的相应设计的6至20个文库。 一组编码蛋白质的基因库。 鉴定与特异性结合配偶体相互作用的蛋白质的方法，该方法包括将蛋白质与蛋白质文库集合的每个文库相结合，观察与该组的某些文库的特异性结合相互作用，并使用观察结果鉴定蛋白质 其与特异性结合配偶体相互作用。 制作随机基因文库的方法。

公开(公告)号：US20060269913A1

公开(公告)日：2006-11-30

申请号：US10518174

申请日：2003-06-13

申请人： Anna Hine ,  David Nagel ,  Marcus Hughes ,  Zhan-Ren Zhang ,  Mohammed Ashraf ,  Andrew Sutherland ,  Albert Santos

发明人： Anna Hine ,  David Nagel ,  Marcus Hughes ,  Zhan-Ren Zhang ,  Mohammed Ashraf ,  Andrew Sutherland ,  Albert Santos

IPC分类号： C40B30/06 ,  C40B40/08

CPC分类号： C12N15/1093 ,  C12N15/67

摘要： The present invention provides a method of producing a DNA library comprising a plurality of DNA sequences of interest, where each DNA sequence of interest has at least two predetermined positions, with at each predetermined position a codon (MAX) selected from a defined group for that position, the codons within a group coding for different amino acids. The method comprising the steps of:—(i) contacting so as to effect hybridisation (a) template DNA (A) comprising said at least two predetermined positions, said template DNA being fully randomised at said at least two predetermined positions (NNN), (b) for each predetermined position, a selection oligonucleotide pool, each selection oligonucleotide (B) within each pool comprising a codon (MAX) selected from the defined group for that predetermined position, and (c) at least one additional oligonucleotide sequence (E) comprising a region (E2) which is non-hybridisable to the template DNA, (ii) ligating the hybridised DNA sequences (B, E), (iii) denaturing the product of step (ii) so as to give a mixed population of said template DNA (A) and said DNA sequences of interest, and (iv) selectively amplifying the DNA sequences of interest. The additional oligonucleotide sequence (E) of step (i) is selected such that after step (ii) the non-hybridisable region (E2) is located externally of the template DNA (A) The invention also provides protein and DNA libraries which can be produced by the method of the invention.

摘要翻译： 本发明提供了一种生产包含多个感兴趣的DNA序列的DNA文库的方法，其中每个目标DNA序列具有至少两个预定位置，在每个预定位置，选自所定义组的密码子（MAX） 位置，编码不同氨基酸的组内的密码子。 所述方法包括以下步骤： - （i）接触以便实现杂交（a）包含所述至少两个预定位置的模板DNA（A），所述模板DNA在所述至少两个预定位置（NNN）完全随机化， （b）对于每个预定位置，选择寡核苷酸库，每个库内的每个选择寡核苷酸（B）包含选自该预定位置的定义基团的密码子（MAX），和（c）至少一个另外的寡核苷酸序列（E ）包含与模板DNA不可杂交的区域（E 2），（ii）连接杂交的DNA序列（B，E），（iii）使步骤（ii）的产物变性以得到混合的群体 的所述模板DNA（A）和所述感兴趣的DNA序列，和（iv）选择性扩增目的DNA序列。 选择步骤（i）的另外的寡核苷酸序列（E）使得在步骤（ii）之后，不可混合区域（E 2）位于模板DNA外部（A）。本发明还提供蛋白质和DNA文库，其可以 通过本发明的方法生产。