用寡聚核苷酸诱导的定位突变法,将人U_1和U_2snRNA基因的5'-端调控区域的一段能与SV_(40)T抗原相结合的DNA删去,造成缺失突变,改变这段DNA核苷酸的排列顺序,造成取代突变。突变株用原位杂交法筛选,由限制性内切酶电泳图谱分析和DNA顺序测定得到证实。突变率约为5％。
Abstract
Using the oligonucleotide directed mutagesis we have deleted a segment from DNA region 5'-flanking of human U1. and U2 snRNA gene which can be bound by SV40 T-antigen. We have also substituted this DNA region with changed nucleotide sequences. The mutants had been screened by in situ colony hybridization. The mutants selected had been characterized by restriction enzyme mapping and DNA sequening. The efficiency of mutagenesis was about 5%.
关键词
U_1U_2基因 /
定位突变 /
缺失 /
取代
{{custom_keyword}} /
Key words
U1 U2 gene /
Mutagenesis /
Deletion /
Substitution
{{custom_keyword}} /
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Charles C S et al.Sciencs,1985;228:291-297．
[2] Donald D S et al.Proc Natl Acad Sci USA 1985;82:2272-2276．
[3] Mark D F et al.Proc Natl Acad Sci USA,1984;81:5662-5666．
[4] Kozak M.Cell,1986;44:283-292．
[5] Norris k et al.Nucl Acids Res 1983;11:5103-5112．
[6] Manitis T et al.Molecular Gloning,Cold Spring Harbor,New York.1982;p122,p250,p315．
[7] Mark J et al.DNA,1984;3:475-488．
[8] Sanger F et al.J Mol Biol,1980;143:161-178．
[9] Maxam.A,Gilbert w.Proc Natl Acad Sci USA,1977;74:560-564．
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
PDF(1327 KB)
