Ribosomal protein L21 (RPL21) plays an important role in ribosome assembly. It is considered to be a major cause for the occurrence of the hypotrichosis simplex (HTS), a type of sustained hair loss from early childhood to adulthood. In this study, the full-length sequence of pig RPL21 gene (GenBank accession number: KU891824) was cloned and identified for the first time. We found it contains a 483-bp open reading frame (ORF) encoding 160 amino acids. It is located in the plus strand of chromosome 11, which spans 2,167 bp from 4,199,792 to 4,201,958. We found RPL21 expression level is closely related to cell proliferation and cell cycle arrest. In the knockdown group, the cell proliferation activity was significantly decreased (P<0.01) and an obvious accumulation of cells at the G2/M phase with a simultaneous up-regulation of p53 and p21 was observed. This likely due to knockdown of RPL21 triggered ribosomal stress, which affected the normal ribosome assembly and caused defective ribosome biogenesis. The unassembled RPs were released consequently from the nucleolus to the nucleoplasm where they can activate p53-dependent cell-cycle responsive factors and led to a G2/M arrest. We expect these results may provide valid information for further study on the pig RPL21 gene and the cause of hypo trichosis simplex.
This study describes an efficient approach to the development of DNA markers for use in distinguishing the Scrophularia species that have been used as useful medicinal crops. In order to distinguish Scrophularia species, DNA sequences of rpl-5 region in mitochondrial DNA of Scrophularia species were analysed for detecting sequence variations, and the PCR-RFLP method was applied for developing practicable DNA marker patterns. Several DNA variations were detected by the sequence comparison of rpl-5 region among Scrophularia species. Genetic relationship analysis of Scrophularia species was carried out based on these DNA variations. DNA variations of rpl-5 region were revealed that it was significantly efficient in genetic relationship analysis of Scrophularia species. In addition, Scrophularia species tested in this study were completely discriminated by four polymorphic genotypes by PCR-RFLP combined with Tsp509 I (^AATT) restriction enzyme. Our results suggested that DNA sequence variations of rpl-5 region were sufficiently useful for genetic relationship analysis of Scrophularia species. Polymorphic genotypes by PCR-RFLP using the Tsp509 I enzyme will be useful for discrimination of Scrophularia species as a practicable DNA markers.