Wood D, Blanchetot A, Jeffreys A J
Nucleic Acids Res. 1982 Nov 25;10(22):7133-44. doi: 10.1093/nar/10.22.7133.
Grey seal skeletal muscle containing high levels of myoglobin was used to prepare poly(A)+ RNA. In vitro translation of this RNA produced a range of polypeptides including myoglobin. cDNA was prepared by reverse transcription of muscle poly(A)+ RNA and cloned into the plasmid pAT 153. 4% of cDNA recombinants were shown to contain myoglobin cDNA inserts. DNA sequence analysis of one clone (pSM 178) which contained a relatively large myoglobin cDNA insert showed an incomplete cDNA comprising the terminal 293 nucleotides of 3' non-translated mRNA sequences. Hybridization experiments using this myoglobin cDNA indicated that seal myoglobin is coded by a single gene which is transcribed to give a 1400 nucleotide mRNA considerably longer than related haemoglobin mRNAs.
含有高水平肌红蛋白的灰海豹骨骼肌被用于制备聚腺苷酸加尾RNA（poly(A)+ RNA）。这种RNA的体外翻译产生了一系列多肽，包括肌红蛋白。通过对肌肉聚腺苷酸加尾RNA进行逆转录制备了互补DNA（cDNA），并将其克隆到质粒pAT 153中。4%的cDNA重组体被证明含有肌红蛋白cDNA插入片段。对一个含有相对较大肌红蛋白cDNA插入片段的克隆（pSM 178）进行的DNA序列分析表明，其cDNA不完整，包含3'非翻译mRNA序列的末端293个核苷酸。使用这种肌红蛋白cDNA进行的杂交实验表明，海豹肌红蛋白由单个基因编码，该基因转录产生一个1400个核苷酸的mRNA，比相关血红蛋白的mRNA长得多。
