Protter A A, Hardman D A, Schilling J W, Miller J, Appleby V, Chen G C, Kirsher S W, McEnroe G, Kane J P
Proc Natl Acad Sci U S A. 1986 Mar;83(5):1467-71. doi: 10.1073/pnas.83.5.1467.
A partial cDNA clone for the B-26 region of apolipoprotein B was isolated from an adult human liver DNA library by screening with an oligonucleotide probe derived from amino-terminal protein sequence obtained from purified B-26 peptide. Antisera against a synthetic 17-residue peptide whose amino acid sequence was encoded by the clone cross-reacts with apolipoproteins B-26, B-100, and B-48, but not with B-74. The nucleotide sequence immediately upstream from the amino terminus of B-26 codes for an apparent signal sequence, implying that the B-26 moiety is in an amino-terminal locus in the B-100 protein. That this sequence represents a 5' end region is further supported by primer extension analysis using a fragment of the cDNA clone and by S1 nuclease protection experiments using the corresponding region in a genomic clone.
通过用从纯化的B - 26肽获得的氨基末端蛋白质序列衍生的寡核苷酸探针筛选，从成人肝脏DNA文库中分离出载脂蛋白B的B - 26区域的部分cDNA克隆。针对由该克隆编码氨基酸序列的合成17肽的抗血清与载脂蛋白B - 26、B - 100和B - 48发生交叉反应，但与B - 74不发生交叉反应。B - 26氨基末端上游紧邻的核苷酸序列编码一个明显的信号序列，这意味着B - 26部分位于B - 100蛋白的氨基末端位点。使用cDNA克隆片段的引物延伸分析以及使用基因组克隆中相应区域的S1核酸酶保护实验进一步支持了该序列代表5'末端区域。
