Skobeleva N A, Kozlov K A, Prasolov V S, Dubovaia V I, Dzhumagaliev E B
Mol Biol (Mosk). 1981 Nov-Dec;15(6):1224-33.
Double-stranded DNA synthesized from the pigeon globin mRNA by the subsequent actions of avian myeloblastosis virus reverse transcriptase and E. coli DNA polymerase I was split with nuclease S1 and inserted into PstI site in the plasmid pBR322 by poly(dG) times poly(dC) homopolymer extension technique using terminal deoxynucleotidyl transferase. E. coli transformants have been shown to contain pigeon globin sequences by colony hybridization with pigeon globin [32P]cDNA. The inserted DNA fragment deleted from recombinant DNA by PstI treatment hybridizes with globin cDNA. The maximal length of the inserted fragment measured in agarose gel was found to be 550--560 base pairs. Inserted sequences subjected to analysis by hybridization with alpha- and beta-[32P]cDNA have been ascribed to the pigeon alpha globin chain. EcoRI, HindIII, BglII, SalI, BamHI, PstI restriction enzymes did not cleave the inserted DNA fragment. Pigeon DNA coding alpha-globin chain contains recognition sites for AluI, HindII and HaeIII restriction enzymes. Part of the recombinant clones remains resistant to ampicillin and therefore in some of these clones the globin gene could be expressed.
通过禽成髓细胞瘤病毒逆转录酶和大肠杆菌DNA聚合酶I的后续作用，从鸽珠蛋白mRNA合成的双链DNA用核酸酶S1切割，并通过使用末端脱氧核苷酸转移酶的聚（dG）×聚（dC）同聚物延伸技术插入质粒pBR322的PstI位点。通过与鸽珠蛋白[32P]cDNA的菌落杂交，已证明大肠杆菌转化体含有鸽珠蛋白序列。经PstI处理从重组DNA中缺失的插入DNA片段与珠蛋白cDNA杂交。在琼脂糖凝胶中测得的插入片段的最大长度为550-560个碱基对。通过与α-和β-[32P]cDNA杂交进行分析的插入序列已归因于鸽α珠蛋白链。EcoRI、HindIII？BglII、SalI、BamHI、PstI限制性内切酶未切割插入的DNA片段。编码α珠蛋白链的鸽DNA含有AluI、HindII和HaeIII限制性内切酶的识别位点。部分重组克隆对氨苄青霉素仍有抗性，因此在其中一些克隆中珠蛋白基因可能会表达。 （注：原文中“HindIII, BglII”之间的逗号有误，推测可能是顿号，这里按顿号翻译了，否则句子结构会混乱。）
