Linnenbach A J, Speicher D W, Marchesi V T, Forget B G
Proc Natl Acad Sci U S A. 1986 Apr;83(8):2397-401. doi: 10.1073/pnas.83.8.2397.
A region of minimal codon degeneracy was selected from the amino acid sequence of the amino-terminal alpha I domain of human erythrocyte spectrin to design a 90-base-pair DNA probe for the screening of a human genomic library. Five complementary oligonucleotides were assembled to form a full-length double-stranded DNA, which was then cloned in an M13 phage vector to generate hybridization probes. Under stringent conditions, a single hybridizing clone was isolated from a total human genomic library. Partial DNA sequence analysis established the 16.8-kilobase-pair isolate as erythrocyte alpha-spectrin by correlation to a known sequence of 131 amino acids. The spectrin 106 amino acid repeat segment is encoded by multiple exons separated by introns of various sizes. Of the 3074 base pairs of DNA sequenced thus far, 12.8% code for amino acids.
从人红细胞血影蛋白氨基末端αI结构域的氨基酸序列中选择密码子简并性最小的区域，设计一个90个碱基对的DNA探针，用于筛选人基因组文库。将五个互补寡核苷酸组装成全长双链DNA，然后克隆到M13噬菌体载体中以产生杂交探针。在严格条件下，从整个人基因组文库中分离出一个单一的杂交克隆。通过与已知的131个氨基酸序列进行比对，部分DNA序列分析确定这个16.8千碱基对的分离片段为红细胞α-血影蛋白。血影蛋白106个氨基酸重复片段由多个被不同大小内含子分隔的外显子编码。在目前已测序的3074个碱基对的DNA中，12.8%编码氨基酸。
