Dame J B, McCutchan T F
J Biol Chem. 1983 Jun 10;258(11):6984-90.
The four ribosomal RNA genes (rDNA units) of the rodent malaria parasite, Plasmodium berghei, were identified and mapped by restriction enzyme analysis and Southern blot hybridization of genomic DNA. Although the four genes share common characteristics, they appear to be internally different from each other in expanse and sequence. One HindIII site near the 3' end of the coding region for the large rRNA is the only restriction site which we have detected that is conserved in all of the genes. The distance between the conserved HindIII site and the coding region for the small rRNA is at least 1-2 kilobases longer in two of the rDNA units than in a third unit. None of the four rDNA units were linked by restriction mapping where about 150 kilobases of the genome were accounted for. The copy number of two of the rDNA units was determined to be approximately 1 per haploid genome by quantitative analysis of cloned (plasmid) DNA versus genomic DNA digests on Southern blots. The other two genes also appear to be present in 1 copy. Restriction analysis confirms both the low copy number and that these genes are not in an easily recognizable tandem array. The low number of rDNA units requires that new copies of the genome produced during intraerythrocytic growth be active in RNA synthesis soon after their replication.
通过对基因组DNA进行限制性酶切分析和Southern杂交,鉴定并定位了啮齿动物疟原虫伯氏疟原虫的四个核糖体RNA基因(rDNA单元)。尽管这四个基因具有共同特征,但它们在长度和序列上似乎内部彼此不同。大rRNA编码区3'端附近的一个HindIII位点是我们检测到的在所有基因中都保守的唯一限制性位点。在两个rDNA单元中,保守的HindIII位点与小rRNA编码区之间的距离比第三个单元至少长1 - 2千碱基。通过限制性酶切图谱分析,在约150千碱基的基因组范围内,这四个rDNA单元均未连锁。通过对Southern印迹上克隆(质粒)DNA与基因组DNA消化产物进行定量分析,确定两个rDNA单元的拷贝数在单倍体基因组中约为1个。另外两个基因似乎也以1个拷贝存在。限制性分析证实了拷贝数低且这些基因并非处于易于识别的串联阵列中。rDNA单元数量少意味着红细胞内生长过程中产生的基因组新拷贝在复制后不久就要活跃于RNA合成。