Development of a novel strategy for mutation-specific cloning using DNA repair enzyme

在这项研究中，我们使用重组MutS蛋白和分离的涉及CEA的突变型和野生型序列对，从具有高微卫星不稳定性（MSI-H）的结直肠癌和配对的正常粘膜构建的cDNA文库之间形成异源双链体。（癌胚抗原相关细胞粘附分子5 ; Hs.220529）3 'UTR，2780 A至G ; 2792-2798（T）7至（T）8（GI：178676 ; CDS 97）。2205）; CCNI（细胞周期蛋白I ; Hs.486360）3' UTR，1800（A）10至（A）9（GI：13436391 ; CDS 544. 1677）; PTMA（胸腺素原，α; Hs.264317）5 'UTR，142 T至C ; CDS，228 T至C（GI：47938172 ; CDS 161. 493); SRI（sorcin ; Hs.489040）CDS 488 A->G，Y159 C（GI：4507206 ; CDS 13. 609)LGALS 3（凝集素，半乳糖苷结合，可溶性，3 ;半乳糖凝集素3 ; Hs.531081）774 A至G，1250 M（GI：37589086 ; CDS 25. 777);RACK 1（GNB 2L 1，鸟嘌呤核苷酸结合蛋白（G蛋白），β多肽2样1 ; Hs.5662）288 t->g，F65 V，304 t->c，V70 A（GI：187701 ; CDS 96. 1049）; UQCRQ（泛喹啉-细胞色素c还原酶，复合物III亚基VII，9.5kDa ; Hs.146602）CDS 227 t->C，F67 S，（GI：34784784 ; CDS 28. 276)还分离了一种在MSI-H肿瘤中高度突变的新型NAT，用于胸苷酸合酶。

In this study, we pulled down heteroduplexes formed between cDNA libraries constructed from a colorectal cancer with high microsatellie instability (MSI-H) and paired normal mucosa using a recombinant MutS protein and isolated paires of mutant and wild type sequences involving ; CEA (carcinoembryonic antigen-related cell adhesion molecule 5 ; Hs.220529) 3'UTR, 2780 A to G ; 2792-2798 (T) 7 to (T) 8 (GI : 178676 ; CDS 97..2205) ; CCNI (Cyclin I ; Hs.486360) 3' UTR, 1800 (A) 10 to (A) 9 (GI : 13436391 ; CDS 544..1677) ; PTMA (prothymosin, alpha ; Hs.264317) 5'UTR, 142 T to C ; CDS, 228 T to C (GI : 47938172 ; CDS 161..493) ; SRI (sorcin ; Hs.489040) CDS 488 A->G, Y159C (GI : 4507206 ; CDS 13..609) ; LGALS3 (lectin, galactoside-binding, soluble, 3 ; galectin 3 ; Hs.531081) 774 A to G, 1250M (GI : 37589086 ; CDS 25..777) ; RACK1 (GNB2L1, guanine nucleotide binding protein (G protein), beta polypeptide 2-like 1 ; Hs.5662) 288t->g, F65V, 304t->c, V70A (GI : 187701 ; CDS 96..1049) ; UQCRQ (ubiquinol-cytochrome c reductase, complex III subunit VII, 9.5kDa ; Hs.146602) CDS 227 t->C, F67S, (GI:34784784 ; CDS 28..276) ; and also isolated a novel NAT highly mutated in MSI-H tumors for thymidylate synthase.