Hayashi M N, Hayashi M
Nucleic Acids Res. 1985 Aug 26;13(16):5937-48. doi: 10.1093/nar/13.16.5937.
The stability of two species of phi X174 polycistronic mRNA in vivo can be altered by mutating sequences existing immediately upstream of a termination site. The wild type phage contains an mRNA stabilizing sequence ((+) sequence), while the same sequence mutated by insertion ((-) sequence) reduces the stability of the mRNAs. These two sequences were cloned at the 3' ends of gene D or gene B of phi X174 in a pBR322 derivative plasmid. The cloned sequences were functional. The (+) sequence stabilized gene B or gene D mRNA; half-lives of these mRNAs were 7 to 8 min. When the (+) sequence is eliminated ((o) sequence) or replaced with the (-) sequence, the half-lives of the mRNA were reduced to about 1 to 2 min. The stabilization of mRNAs caused an increased production of these proteins.
通过突变位于终止位点紧邻上游的序列,可改变两种X174噬菌体多顺反子mRNA在体内的稳定性。野生型噬菌体含有一个mRNA稳定序列((+)序列),而同一序列经插入突变后((-)序列)会降低mRNA的稳定性。将这两个序列克隆到pBR322衍生质粒中X174噬菌体基因D或基因B的3'端。克隆的序列具有功能。(+)序列使基因B或基因D的mRNA稳定;这些mRNA的半衰期为7至8分钟。当(+)序列缺失((o)序列)或被(-)序列取代时,mRNA的半衰期缩短至约1至2分钟。mRNA的稳定导致这些蛋白质的产量增加。