Genetic improvement of meat production traits has always been the primary goal of pig breeding.Geographical isolation,natural and artificial selection led to significant differences in the phenotypes of meat productio...Genetic improvement of meat production traits has always been the primary goal of pig breeding.Geographical isolation,natural and artificial selection led to significant differences in the phenotypes of meat production traits between Chinese local pigs and Western commercial pigs.Comparative genomics and transcriptomics analysis provided powerful tools to identify genetic variants and genes associated with skeletal muscle growth.However,the number of available genetic variants and genes are still limited.In this study,a comprehensive comparison of transcriptomes showed that ribosomal protein S27-like(RPS27L)gene was highly expressed in skeletal muscle and up-regulated in Chinese local pigs when compared with Western commercial pigs.Functional analysis revealed that overexpression of RPS27L promoted myoblast proliferation and repressed differentiation in pig skeletal muscle cells.Conversely,the knockdown of RPS27L led to the inhibition of myoblast proliferation and the promotion of differentiation.Notably,a 13-bp insertion-deletion(InDel)mutation was identified within the RPS27L promoter,inserted in Chinese local breeds and predominantly deleted in Western commercial breeds.Luciferase reporter assay suggested this InDel modulated RPS27L expression by influencing transcription factor 3(TCF3)and myogenic differentiation antigen(MYOD)binding to the promoter.Furthermore,a positive correlation was observed between RPS27L expression and backfat thickness.Association studies demonstrated this InDel was significantly associated with the body weight of pigs at the age of 240 d.Together,our results suggested that RPS27L was a regulator of skeletal muscle development and growth,and was a candidate marker for improving meat production traits in pigs.This study not only provided a biomarker for animal breeding,but also was helpful for understanding skeletal muscle development and muscular disease in humans.展开更多
The DNA replication stress(RS)response is crucial for maintaining cellular homeostasis and promoting physiological longevity.However,the mechanisms by which long-lived species,such as bats,regulate RS to maintain geno...The DNA replication stress(RS)response is crucial for maintaining cellular homeostasis and promoting physiological longevity.However,the mechanisms by which long-lived species,such as bats,regulate RS to maintain genomic stability remain unclear.Also,recent studies have uncovered noncanonical roles of ribosome-associated factors in maintaining genomic stability.In this study,somatic skin fibroblasts from the long-lived big-footed bat(Myotis pilosus)were examined,with results showing that bat cells exhibited enhanced RS tolerance compared to mouse cells.Comparative transcriptome analysis under RS conditions revealed pronounced species-specific transcriptional differences,including robust up-regulation of ribosome biogenesis genes in bat cells and a markedly reduced activation of the P53 signaling pathway.These features emphasize a distinct homeostatic strategy in bat cells.Nuclear fragile X mental retardation-interacting protein 1(Nufip1),a ribosome-associated factor highly expressed in bat fibroblasts,was identified as a potential integrator of ribosomal and P53 signaling via its association with ribosomal protein S27-like(Rps27l).These findings provide direct cellular and molecular evidence for a noncanonical RS response in bats,highlighting a deeper understanding of the biological characteristics and genomic maintenance mechanisms of long-lived species.展开更多
Through mRNA extract, RT and a series of PCR, phage antibody libraries were constructed from rP27kip1-immunized and non-immunized mice. After only one round of selection with rP27kip1, clones from each library were ch...Through mRNA extract, RT and a series of PCR, phage antibody libraries were constructed from rP27kip1-immunized and non-immunized mice. After only one round of selection with rP27kip1, clones from each library were chosen randomly and digested by Tag I and Hinf I. 11 of 64 clones from the immunized animal had consistent restriction pattern, while none of the 64 clones from the non-immunized animal had, except that one had the same fragments pattern as that of the 11 clones. The 12 fragments were expressed in E. coli BL2l(DE3)/pET-28b( + ) system. ELISA showed that some of the fragments could bind to rP27kip1 specifically. All these results implied that specific antibody can be obtained by genetic engineering without hybridoma or many rounds of growth and panning selection.展开更多
基金supported by the Sustainable Development Special Project from Shenzhen,China(KCXFZ20201221173213037)the National Natural Science Foundation of China(32172697 and U23A20229)+1 种基金the Guangdong Provincial Natural Science Foundation(2021A1515011336)the Agricultural Science and Technology Innovation Program,China(CAASZDRW202406)。
文摘Genetic improvement of meat production traits has always been the primary goal of pig breeding.Geographical isolation,natural and artificial selection led to significant differences in the phenotypes of meat production traits between Chinese local pigs and Western commercial pigs.Comparative genomics and transcriptomics analysis provided powerful tools to identify genetic variants and genes associated with skeletal muscle growth.However,the number of available genetic variants and genes are still limited.In this study,a comprehensive comparison of transcriptomes showed that ribosomal protein S27-like(RPS27L)gene was highly expressed in skeletal muscle and up-regulated in Chinese local pigs when compared with Western commercial pigs.Functional analysis revealed that overexpression of RPS27L promoted myoblast proliferation and repressed differentiation in pig skeletal muscle cells.Conversely,the knockdown of RPS27L led to the inhibition of myoblast proliferation and the promotion of differentiation.Notably,a 13-bp insertion-deletion(InDel)mutation was identified within the RPS27L promoter,inserted in Chinese local breeds and predominantly deleted in Western commercial breeds.Luciferase reporter assay suggested this InDel modulated RPS27L expression by influencing transcription factor 3(TCF3)and myogenic differentiation antigen(MYOD)binding to the promoter.Furthermore,a positive correlation was observed between RPS27L expression and backfat thickness.Association studies demonstrated this InDel was significantly associated with the body weight of pigs at the age of 240 d.Together,our results suggested that RPS27L was a regulator of skeletal muscle development and growth,and was a candidate marker for improving meat production traits in pigs.This study not only provided a biomarker for animal breeding,but also was helpful for understanding skeletal muscle development and muscular disease in humans.
基金supported by the Applied Basic Research Programs of Science and Technology Commission Foundation of Yunnan Province(202401AT070186 to K.Q.L.,202201AS070044 to B.Z.)Yunnan Province(202305AH340006 to B.Z.)Kunming Science and Technology Bureau(2022SCP007 to B.Z.)。
文摘The DNA replication stress(RS)response is crucial for maintaining cellular homeostasis and promoting physiological longevity.However,the mechanisms by which long-lived species,such as bats,regulate RS to maintain genomic stability remain unclear.Also,recent studies have uncovered noncanonical roles of ribosome-associated factors in maintaining genomic stability.In this study,somatic skin fibroblasts from the long-lived big-footed bat(Myotis pilosus)were examined,with results showing that bat cells exhibited enhanced RS tolerance compared to mouse cells.Comparative transcriptome analysis under RS conditions revealed pronounced species-specific transcriptional differences,including robust up-regulation of ribosome biogenesis genes in bat cells and a markedly reduced activation of the P53 signaling pathway.These features emphasize a distinct homeostatic strategy in bat cells.Nuclear fragile X mental retardation-interacting protein 1(Nufip1),a ribosome-associated factor highly expressed in bat fibroblasts,was identified as a potential integrator of ribosomal and P53 signaling via its association with ribosomal protein S27-like(Rps27l).These findings provide direct cellular and molecular evidence for a noncanonical RS response in bats,highlighting a deeper understanding of the biological characteristics and genomic maintenance mechanisms of long-lived species.
文摘以人外周血为研究对象,研究照射后RPS27L基因的时间效应和剂量效应以及本底水平,探究其作为辐射损伤早期生物标志物的可行性。采集3名健康成人外周血,利用γ射线对其进行照射至吸收剂量分别为0、0.5、1、2、4、6和10 Gy,受照后37℃培养2、4、8、12和24 h,利用实时荧光定量PCR检测RPS27L基因的表达变化,并分析在37个健康人个体中的本底水平。结果显示,受照后的4、8、12和24 h RPS27L基因表达变化显著,而且具有很好的剂量依赖关系;在不同年龄和性别的个体中RPS27L本底水平相对稳定。因此,RPS27L基因具有作为新的辐射损伤标志物的潜力。
文摘Through mRNA extract, RT and a series of PCR, phage antibody libraries were constructed from rP27kip1-immunized and non-immunized mice. After only one round of selection with rP27kip1, clones from each library were chosen randomly and digested by Tag I and Hinf I. 11 of 64 clones from the immunized animal had consistent restriction pattern, while none of the 64 clones from the non-immunized animal had, except that one had the same fragments pattern as that of the 11 clones. The 12 fragments were expressed in E. coli BL2l(DE3)/pET-28b( + ) system. ELISA showed that some of the fragments could bind to rP27kip1 specifically. All these results implied that specific antibody can be obtained by genetic engineering without hybridoma or many rounds of growth and panning selection.
