Tandeau de Marsac N, Mazel D, Bryant D A, Houmard J
Nucleic Acids Res. 1985 Oct 25;13(20):7223-36. doi: 10.1093/nar/13.20.7223.
Since the gas vesicle protein (GVP) is highly conserved among the different gas-vacuolate prokaryotes, a 29-mer oligonucleotide corresponding to a portion of the Anabaena flos-aquae GVP gene was synthesized and used to isolate the GVP structural gene from Calothrix PCC 7601 (= Fremyella diplosiphon). Gas vacuole production in this filamentous cyanobacterium is restricted to hormogonia which occur at a specific stage during the developmental cell cycle. The GVP gene (gvpA) was localized on a 709 bp HindIII-HincII fragment. Nucleotide sequence analysis revealed a 213 bp open reading frame whose deduced amino-acid sequence shows a very high homology with that of the Anabaena flos-aquae GVP. Assuming that the first methionine residue is proteolytically processed, the molecular mass of the Calothrix GVP is 7375 daltons. Sequences resembling the Escherichia coli consensus promoter were found upstream from the gvpA gene. The initiator codon of the gvpA gene is preceded by a polypurine sequence assumed to be the ribosome binding site. Southern hybridizations with a probe specific for the gvpA gene indicated that this gene is not plasmid-borne, and that another homologous gene is present in the Calothrix genome.
由于气体囊泡蛋白(GVP)在不同的产气泡原核生物中高度保守,因此合成了一段与水华鱼腥藻GVP基因部分序列对应的29聚体寡核苷酸,并用于从鞘丝藻PCC 7601(=双歧鱼腥藻)中分离GVP结构基因。在这种丝状蓝细菌中,气体囊泡的产生仅限于在发育细胞周期的特定阶段出现的藻殖段。GVP基因(gvpA)定位在一个709 bp的HindIII-HincII片段上。核苷酸序列分析揭示了一个213 bp的开放阅读框,其推导的氨基酸序列与水华鱼腥藻GVP的氨基酸序列具有非常高的同源性。假设第一个甲硫氨酸残基经过蛋白水解加工,鞘丝藻GVP的分子量为7375道尔顿。在gvpA基因上游发现了类似于大肠杆菌共有启动子的序列。gvpA基因的起始密码子之前有一个聚嘌呤序列,假定为核糖体结合位点。用gvpA基因特异性探针进行的Southern杂交表明,该基因不是质粒携带的,并且鞘丝藻基因组中存在另一个同源基因。