Steppuhn J, Hermans J, Nechushtai R, Ljungberg U, Thümmler F, Lottspeich F, Herrmann R G
Botanisches Institut der Ludwig-Maximilians-Universität, München, FRG.
FEBS Lett. 1988 Sep 12;237(1-2):218-24. doi: 10.1016/0014-5793(88)80205-9.
Using lambda gt11 expression cloning and immunoscreening, cDNA-containing recombinant phages for subunits IV and V of the photosystem I reaction center were isolated, sequenced and used to probe Northern blots of polyadenylated RNA prepared from spinach seedlings. The mRNA sizes for both components are approximately 1000 and 850 nucleotides, respectively. The 968 nucleotide cDNA sequence and derived amino acid sequence for subunit IV predict a single open reading frame of 231 amino acid residues (25.4 kDa). Comparison with a 13-residue N-terminal amino acid sequence determined for subunit IV suggests a mature protein of 17.3 kDa (154 residues) and a transit sequence of 77 amino acids (8.1 kDa). The corresponding data for subunit V are 677 bp (cDNA), 167 residues for the precursor protein (18.2 kDa), 98 residues for the mature polypeptide (10.8 kDa) and 69 residues for the transit peptide (7.4 kDa). Secondary structure predictions indicate that both proteins possess greatly different transit sequences and that none is membrane-spanning.
利用λgt11表达克隆和免疫筛选技术,分离出了编码光系统I反应中心亚基IV和亚基V的含cDNA重组噬菌体,对其进行测序并用于探测从菠菜幼苗制备的聚腺苷酸化RNA的Northern杂交印迹。这两种组分的mRNA大小分别约为1000和850个核苷酸。亚基IV的968个核苷酸cDNA序列及其推导的氨基酸序列预测有一个231个氨基酸残基(25.4 kDa)的单一开放阅读框。与为亚基IV测定的13个残基的N端氨基酸序列比较表明,成熟蛋白为17.3 kDa(154个残基),转运序列为77个氨基酸(8.1 kDa)。亚基V的相应数据为677 bp(cDNA),前体蛋白167个残基(18.2 kDa),成熟多肽98个残基(10.8 kDa),转运肽69个残基(7.4 kDa)。二级结构预测表明,这两种蛋白质具有非常不同的转运序列,且均无跨膜结构。