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从莱茵衣藻中分离和鉴定编码光系统 I 的 17.9 和 8.1 kDa 亚基的 cDNA 克隆。

Isolation and characterization of cDNA clones encoding the 17.9 and 8.1 kDa subunits of Photosystem I from Chlamydomonas reinhardtii.

机构信息

Departments of Molecular Biology and Plant Biology, University of Geneva, Geneva, Switzerland.

出版信息

Plant Mol Biol. 1989 Apr;12(4):463-74. doi: 10.1007/BF00017585.

Abstract

cDNA clones encoding two Photosystem I subunits of Chlamydomonas reinhardtii with apparent molecular masses of 18 and 11 kDa (thylakoid polypeptides 21 and 30; P21 and P30 respectively) were isolated using oligonucleotides, the sequences of which were deduced from the N-terminal amino acid sequences of the proteins. The cDNAs were sequenced and used to probe Southern and Northern blots. The Southern blot analysis indicates that both proteins are encoded by single-copy genes. The mRNA sizes of the two components are 1400 and 740 nucleotides, respectively. Comparison between the open reading frames of the cDNAs and the N-terminal amino acid sequences of the proteins indicates that the molecular masses of the mature proteins are 17.9 (P21) and 8.1 kDa (P30). Analysis of the deduced protein sequences predicts that both subunits are extrinsic membrane proteins with net positive charges. The amino acid sequences of the transit peptides suggest that P21 and P30 are routed towards the lumenal and stromal sides of the thylakoid membranes, respectively.

摘要

使用寡核苷酸分离了编码莱茵衣藻两个光系统 I 亚基的 cDNA 克隆,它们的表观分子量分别为 18 和 11 kDa(类囊体多肽 21 和 30;分别为 P21 和 P30)。这些寡核苷酸的序列是根据蛋白质的 N 末端氨基酸序列推导出来的。cDNA 进行了测序,并用于探测 Southern 和 Northern 印迹。Southern 印迹分析表明,这两种蛋白质均由单拷贝基因编码。两个组件的 mRNA 大小分别为 1400 和 740 个核苷酸。cDNA 的开放阅读框与蛋白质的 N 末端氨基酸序列的比较表明,成熟蛋白的分子量分别为 17.9(P21)和 8.1 kDa(P30)。对推导的蛋白质序列的分析表明,两个亚基都是带有净正电荷的外在膜蛋白。转运肽的氨基酸序列表明,P21 和 P30 分别被导向类囊体膜的腔和基质侧。

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