Adam R D
University of Arizona Health Sciences Center, Section of Infectious Disease and Microbiology & Immunology, Tucson, AZ 85724.
Nucleic Acids Res. 1992 Jun 25;20(12):3057-61. doi: 10.1093/nar/20.12.3057.
Giardia lamblia trophozoites contain at least five sets of chromosomes that have been categorized by chromosome-specific probes. Pulsed field separations of G. lamblia chromosomes also demonstrated minor bands in some isolates which stained less intensely with ethidium than the major chromosomal bands. Two of the minor bands of the E11 clone of the ISR isolate, MBa and MBb, were similar to each other and to chromosomal band I by hybridization to total chromosomal DNA and by hybridization of specific probes. In order to determine the extent of this similarity, I have developed a panel of probes for many of the Pacl restriction fragments and have shown that most of the Pacl and Notl fragments found in MBa are also present in MBb. The differences are found in both telomeric regions. At one end, MBb contains a 300 kb region not found in MBa. At the other end of MBb is a 160 kb region containing the rDNA repeats which is bounded on one end by the telomeric repeat and on the other by sites for multiple enzymes that do not digest the rDNA repeats. The corresponding region of MBa is 23 kb in size. The size difference is consistent with the eightfold greater number of rDNA repeats in MBb than MBa and suggests that 30% of the size difference is accounted for by different numbers of copies of the rDNA repeat. MBa of another ISR clone (ISR G5) is 150 kb larger in size than MBa of ISR E11. The data suggest that MBa and MBb are homologous chromosomes of different sizes and that a portion of the size difference is accounted for by different copy numbers of the rDNA repeat.
蓝氏贾第鞭毛虫滋养体含有至少五组已通过染色体特异性探针分类的染色体。蓝氏贾第鞭毛虫染色体的脉冲场分离也表明,在一些分离株中存在一些较细的条带,这些条带用溴化乙锭染色时比主要染色体条带的染色强度弱。通过与总染色体DNA杂交以及特定探针杂交,ISR分离株E11克隆的两条较细条带MBa和MBb彼此相似,且与染色体条带I相似。为了确定这种相似性的程度,我针对许多Pacl限制性片段开发了一组探针,并表明在MBa中发现的大多数Pacl和Notl片段在MBb中也存在。差异存在于两个端粒区域。在一端,MBb包含一个在MBa中未发现的300 kb区域。在MBb的另一端是一个160 kb的区域,包含rDNA重复序列,该区域一端由端粒重复序列界定,另一端由多种不消化rDNA重复序列的酶的位点界定。MBa的相应区域大小为23 kb。大小差异与MBb中rDNA重复序列的数量比MBa多八倍一致,这表明30%的大小差异是由rDNA重复序列的不同拷贝数造成的。另一个ISR克隆(ISR G5)的MBa比ISR E11的MBa大150 kb。数据表明,MBa和MBb是不同大小的同源染色体,且部分大小差异是由rDNA重复序列的不同拷贝数造成的。