Chiaruttini C, Expert-Bezançon A, Hayes D, Ehresmann B
Nucleic Acids Res. 1982 Dec 11;10(23):7657-76. doi: 10.1093/nar/10.23.7657.
1-ethyl-3-dimethyl aminopropylcarbodiimide (EDC) was used to cross-link 30S ribosomal proteins to 16S rRNA within the E. coli 3OS ribosomal subunit. Covalently linked complexes containing 30S proteins and 16S rRNA, isolated by sedimentation of dissociated crosslinked 30S subunits through SDS containing sucrose gradients, were digested with RNase T1, and the resulting oligonucleotide-protein complexes were fractionated on SDS containing polyacrylamide gels. Eluted complexes containing 30S proteins S9 and S12 linked to oligonucleotides were obtained in pure form. Oligonucleotide 5'terminal labelling was successful in the case of S12 containing but not of the S9 containing complex and led to identification of the S12 bound oligonucleotide as CAACUCG which is located at positions 1316-1322 in the 16S rRNA sequence. Protein S12 is crosslinked to the terminal G of this heptanucleotide.
1-乙基-3-二甲基氨基丙基碳二亚胺（EDC）用于将大肠杆菌30S核糖体亚基中的30S核糖体蛋白与16S rRNA交联。通过在含蔗糖梯度的SDS中对解离的交联30S亚基进行沉降分离得到的含有30S蛋白和16S rRNA的共价连接复合物，用核糖核酸酶T1消化，然后将所得的寡核苷酸-蛋白质复合物在含SDS的聚丙烯酰胺凝胶上进行分级分离。得到了以纯形式存在的含有与寡核苷酸相连的30S蛋白S9和S12的洗脱复合物。在含S12的复合物中寡核苷酸5'末端标记成功，但在含S9的复合物中未成功，这使得与S12结合的寡核苷酸被鉴定为CAACUCG，其位于16S rRNA序列的1316 - 1322位。蛋白S12与这个七核苷酸的末端G交联。
