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恶性疟原虫主要RNA种类的初步鉴定

Preliminary characterization of the major RNA species from Plasmodium falciparum.

作者信息

Vezza A C, Trager W

出版信息

Mol Biochem Parasitol. 1981 Dec;4(3-4):149-62. doi: 10.1016/0166-6851(81)90014-1.

Abstract

Polyacrylamide gel electrophoresis of the total RNA extracted from cultures of Plasmodium falciparum resolved two major RNA components with estimated molecular weights of 1.3 X 10(6) and 0.72 - 0.74 X 10(6). Oligo(dT)-cellulose column chromatography of the total cellular RNA indicates that more than 90% of the total radiolabeled RNA lacks substantial poly(A) sequences (poly(A)-). Resolution of the poly(A)- fraction on polyacrylamide or agarose gels indicates that the majority of the poly(A)- RNAs are the 1.3 X 10(6) and 0.72 - 0.74 X 10(6) species. This observation was confirmed by two-dimensional oligonucleotide fingerprint analysis of the total cellular RNA as well as the poly(A)- RNA components. Oligonucleotide fingerprint analysis confirmed that the small RNA species is not a breakdown or cleavage product of the larger RNA component. Base ratio analysis of the large and small RNA species indicate that they are typically protozoan type with a low guanine + cytosine (G + C) content. The findings that these two RNA species (i) are the major cellular RNA species, (ii) lack substantial poly(A) sequences, (iii) have estimated molecular weights similar to the ribosomal RNAs obtained from other protozoa, (iv) have a low G + C content (approximately equal to 35 - 37%), and (v) are distinct from one another, indicates that the 1.3 X 10(6) and 0.72 - 0.74 X 10(6) RNA components obtained from P. falciparum cultures are the major ribosomal RNAs.

摘要

对从恶性疟原虫培养物中提取的总RNA进行聚丙烯酰胺凝胶电泳,分离出了两个主要的RNA组分,其估计分子量分别为1.3×10⁶和0.72 - 0.74×10⁶。对总细胞RNA进行寡聚(dT)-纤维素柱层析表明,超过90%的总放射性标记RNA缺乏大量的多聚(A)序列(多聚(A)⁻)。在聚丙烯酰胺或琼脂糖凝胶上对多聚(A)⁻组分进行分离表明,大多数多聚(A)⁻RNA是1.3×10⁶和0.72 - 0.74×10⁶的种类。通过对总细胞RNA以及多聚(A)⁻RNA组分进行二维寡核苷酸指纹分析,证实了这一观察结果。寡核苷酸指纹分析证实,小RNA种类不是大RNA组分的降解或切割产物。对大、小RNA种类的碱基比率分析表明,它们通常是原生动物类型,鸟嘌呤 + 胞嘧啶(G + C)含量低。这两个RNA种类的以下发现:(i)是主要的细胞RNA种类,(ii)缺乏大量的多聚(A)序列,(iii)估计分子量与从其他原生动物获得的核糖体RNA相似,(iv)G + C含量低(约等于35 - 37%),以及(v)彼此不同,表明从恶性疟原虫培养物中获得的1.3×10⁶和0.72 - 0.74×10⁶RNA组分是主要的核糖体RNA。

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