Lau R H, Alvarado-Urbina G, Lau P C
Gene. 1987;52(1):21-9. doi: 10.1016/0378-1119(87)90391-x.
The cloning and nucleotide sequence determination of the Anacystis nidulans R2 phycocyanin (PC) alpha-subunit gene are described. A 3.0-kb PstI fragment of Anacystis nidulans R2 genomic DNA cloned in plasmid pUC8 was found to hybridize with a heptadecameric oligodeoxynucleotide probe. Sequencing using synthetic primers revealed the presence of the PC alpha-subunit gene and the 3' proximal end of the beta-subunit gene. The alpha-gene is separated from the upstream beta-gene by a spacer length of 51 bp. The deduced amino acid (aa) sequence of the alpha-subunit protein is identical, except for 5 aa, to that of A. nidulans 6301 and is highly homologous (77%) to that reported for Agmenellum quadruplicatum PR6. The 16-kDa alpha-subunit protein, detected by immunoadsorption, was fortuitously expressed in Escherichia coli from the lacZ promoter of the cloning vehicle pUC8.
描述了集胞藻6301 R2藻蓝蛋白（PC）α亚基基因的克隆及核苷酸序列测定。发现克隆于质粒pUC8中的集胞藻6301 R2基因组DNA的一个3.0 kb PstI片段与一个十七聚体寡脱氧核苷酸探针杂交。使用合成引物进行测序揭示了PC α亚基基因和β亚基基因3'近端的存在。α基因与上游的β基因被一个51 bp的间隔区隔开。α亚基蛋白推导的氨基酸（aa）序列，除了5个氨基酸外，与集胞藻6301的相同，并且与报道的四聚小球藻PR6的高度同源（77%）。通过免疫吸附检测到的16 kDa α亚基蛋白，偶然地从克隆载体pUC8的lacZ启动子在大肠杆菌中表达。
