Yeast artificial chromosomes from a human genomic

px215467

Yeast artificial chromosomes from the human genome sequence fragment of the expression of direct separation of Study

gNS, CollinsFS-TheHuntfortheneurofibromams ~ gene. ArchDermato1.127: 1705 ~ l707. 11GrodenJ, ThllverisA,[link widoczny dla zalogowanych], SamowitzW, eta1. Identi [icationandch ~ racterirationotthefamilialaderv)-matOUSpolyposiscoilgene. Cel1.1991, 66:589 ~ 600PreliminarystudyondirectisolationofthehumangenomiccodingsequencefromyeastartificialchromosomeShiShaolin, Yulong, LiuMengmin, DengYu, ZhangMIn, GongRuomu. HanShunsheng, XuYuefang, ZhaoShouyan, TanJiazhen (InstituteofGenetics) Al ~ tractThisisthefirstreportinChinaonthemethodofdirectscreeningofCO-dingsequencesfromhumangenomieYACclones. YAC27D08locatedonchromOSOme13q14 · 3wasusedasaprobeforscreeninghumanbraincDNAlibrary, 52possibleposi- 4 Shi Shaolin , etc.; from yeast artificial stained fragment of the human genome pensions direct expression of the order of the initial separation of 369tivecloneswereselectedfrom1 × 10'cDNAclones. Amongthem, 21possibleclonesweredistinguishedfromtheother31falsepositiveclonesbyhybridizationtheSouthernblotofPCR-amplifiedDroductsofthesecloneswithYAC271D08. ThreecandidatecDNAcloneswereidentified (fromthese21positivedones) byhybridizationusinghu-mangenomicDNA, yeastgenomicDNAandhumanrDNAasprobes · ThesethreewerefurtherhybridizedwithYAC27D] 08andother10YACs1ocatedelsewlhere. Thegenera ~ a edcDNAswereconfirmedasYAC27D. 8 a specificandthusdesignatedascfdl3-1 (2.0 kb), cfd13-2 (1.1kb), cfdl3 - 3 (3.8kb). ByNorthernhybridizattlonitwasshownthatthetranscriptscorrespondingtocfdl3 - 1andcfdl3-2wereofthelengths9 · 0kband7 · 0kb. respectively. Whereas, cfdl3. -3showed6bandswhichwereof12 · 0,3 · 0,2 · 2,1 · 5,1 · 0and0.6kbinlength. KeywordsYAC; humangenomelcDNFAclone; directscreening