利用Hoechst 33258荧光染色法检测紫外线B(UVB)辐射诱导HaCaT细胞凋亡率。结果表明,扇贝多肽(Polypeptide from Chlamys farreri,PCF)可以剂量依赖性抑制UVB诱导的HaCaT细胞凋亡;表皮生长因子受体(EGFR)抑制剂AG1478能明显抑制UVB诱导的...利用Hoechst 33258荧光染色法检测紫外线B(UVB)辐射诱导HaCaT细胞凋亡率。结果表明,扇贝多肽(Polypeptide from Chlamys farreri,PCF)可以剂量依赖性抑制UVB诱导的HaCaT细胞凋亡;表皮生长因子受体(EGFR)抑制剂AG1478能明显抑制UVB诱导的HaCaT细胞凋亡。采用3’-RACE法构建EGFR的cDNA片段,克隆测序检测突变位点。结果表明,UVB照射后EGFR发生碱基突变A→G,A→G,T→C,G→A,G→A;预加入5.69mmol/L的PCF,产生部分抗突变作用,1,2,3突变位点处未发生碱基的突变。展开更多
Based on the research of juvenile (2, 3, 4 months) growth and survival of three populations of two different geographic areas in Chlamys farreri from Russian and China and their F1 hybrids derived from Chinese cultura...Based on the research of juvenile (2, 3, 4 months) growth and survival of three populations of two different geographic areas in Chlamys farreri from Russian and China and their F1 hybrids derived from Chinese cultural population (CC) (?) × Russian population (RW) (?) , Chinese wild population (CW) (?) × Russian population (RW) (?), Russian population (RW) (?) × Chinese wild population (CW) (?) , the study of the medium-term (6, 8, 10, 12 months) growth and development of Chlamys farreri was carried out. The four determined results indicated that there existed different extent heterosis (3% -52%) for the growth in three types of F1 hybrids, and the offspring derived from CC(?) ×R(?) had a stronger heterosis among the crosses at the medium-term; the uptrend among traits are wet weight >shell width>shell length> shell height, Chinese cultural population could be recognized as excellent parent, and seasonal variations influence very much on the daily increment and growth rate of each trait of Chlamys farreri and it is only able to survive and could barely grow in winter (6-8 months), but grows fast in temperate season (10-12 months).展开更多
Toll-like receptor(TLR) signaling pathway plays a pivotal role in the innate immune system. Studies on TLR signaling pathway genes in Zhikong scallop(Chlamys farreri) have mainly focused on sequence analysis and expre...Toll-like receptor(TLR) signaling pathway plays a pivotal role in the innate immune system. Studies on TLR signaling pathway genes in Zhikong scallop(Chlamys farreri) have mainly focused on sequence analysis and expression profiling, no research has been carried out on their localization. The chromosomal position of TLR signaling pathway genes can be valuable for assemblying scallop genome and analysizing gene regulatory networks. In the present study, five key TLR signaling pathway genes(Cf TLR, Cf Myd88, Cf TRAF6, Cf NFκB, and Cf IκB) containing bacterial artificial chromosomes(BACs) were isolated and physically mapped through fluorescence in situ hybridization on five non-homologous chromosome pairs, showing a similar distribution to another five model species. The isolation and mapping of these key immune genes of C. farreri will aid to the research on innate immunity, assignment of interested genes to chromosomes, and integration of physical, linkage and cytogenetic maps of this species.展开更多
This study applied an optimized one-step 2b-RAD library construction strategy and performed simplified genome sequencing of 539 individuals from three continuously selected Chlamys farreri populations. SNP screening w...This study applied an optimized one-step 2b-RAD library construction strategy and performed simplified genome sequencing of 539 individuals from three continuously selected Chlamys farreri populations. SNP screening was performed using RAD typing software and population genetic parameters for the continuously selected populations from three generations(G1, G2, G3) were determined. The results showed that the optimized one-step 2b-RAD library construction strategy greatly simplified the experimental process, making it suitable for efficiently constructing a large number of libraries. A total of 18450 SNP markers were identified, which evenly distributed throughout the genome. Population genetic analysis of these three generations showed that the mean value of observed heterozygosity was 0.275 ± 0.177, 0.272 ± 0.181 and 0.275 ± 0.166, respectively. Meanwhile, the mean value of expected heterozygosity was 0.275 ± 0.141, 0.274 ± 0.145 and 0.280 ± 0.133, respectively. The Wright's fixation index(F) was 0.04291, 0.04976 and 0.06685, respectively. Markers deviated from Hardy–Weinberg equilibrium accounted for 10.34%, 12.64%, and 23.11%, and the Shannon diversity index was 0.0999 ± 0.0404, 0.0921 ± 0.0388 and 0.0733 ± 0.0308. FIS(also known as the inbreeding coefficient) of the three populations was 0.0256, 0.0323 and 0.0468, respectively. We suggested that the 2b-RAD method is well suited to population genetic studies of aquacultured organisms. Moreover, our results indicated that the continuous selection affected the population genetic structure of the cultured Penglai-Red scallop, but the change was not significant; therefore, population selection should continue.展开更多
DAX1, a member of nuclear receptor superfamily, has a function in the sex determination and gonadal differentiation of several vertebrate species. However, little information about DAX1 of invertebrates is available. ...DAX1, a member of nuclear receptor superfamily, has a function in the sex determination and gonadal differentiation of several vertebrate species. However, little information about DAX1 of invertebrates is available. Here we cloned a homolog of scallop(Chlamys farreri Jones and Preston 1904) dax1, Cf-dax1, and determined its expression characteristics at m RNA and protein levels. The c DNA sequence of Cf-dax1 was 2093 bp in length, including 1404 bp open reading frame(ORF) encoding 467 amino acids. Unlike those of vertebrates, no conserved LXXLL-related motif was found in the putative DNA binding region of Cf-DAX1. Fluorescence in situ hybridization showed that Cf-dax1 located on the short arm of a pair of subtelocentric chromosomes. Tissue distribution analysis using semi-quantitative RT-PCR revealed that Cf-dax1 expressed widely in adult scallop tissues, with the highest expression level found in adductor muscle, moderate level in mantle, gill and testis, and low level in kidney, ovary and hepatopancreas. The result of quantitative real-time PCR indicated that the expression of Cf-dax1 was significantly higher(P<0.05) in testis than in ovary at the same stage, showing a sex-dimorphic expression pattern. Furthermore, immunohistochemical detection found that Cf-DAX1 mainly located in spermatogonia and spermatocytes of testis and in oogonia and oocytes of ovary, implying that DAX1 may involve in gametogenesis of bivalves.展开更多
文摘利用Hoechst 33258荧光染色法检测紫外线B(UVB)辐射诱导HaCaT细胞凋亡率。结果表明,扇贝多肽(Polypeptide from Chlamys farreri,PCF)可以剂量依赖性抑制UVB诱导的HaCaT细胞凋亡;表皮生长因子受体(EGFR)抑制剂AG1478能明显抑制UVB诱导的HaCaT细胞凋亡。采用3’-RACE法构建EGFR的cDNA片段,克隆测序检测突变位点。结果表明,UVB照射后EGFR发生碱基突变A→G,A→G,T→C,G→A,G→A;预加入5.69mmol/L的PCF,产生部分抗突变作用,1,2,3突变位点处未发生碱基的突变。
基金This study is contribution No. G199901209 of 973 from the Chinese Basic Research Project and Chinese Postdoctor Found
文摘Based on the research of juvenile (2, 3, 4 months) growth and survival of three populations of two different geographic areas in Chlamys farreri from Russian and China and their F1 hybrids derived from Chinese cultural population (CC) (?) × Russian population (RW) (?) , Chinese wild population (CW) (?) × Russian population (RW) (?), Russian population (RW) (?) × Chinese wild population (CW) (?) , the study of the medium-term (6, 8, 10, 12 months) growth and development of Chlamys farreri was carried out. The four determined results indicated that there existed different extent heterosis (3% -52%) for the growth in three types of F1 hybrids, and the offspring derived from CC(?) ×R(?) had a stronger heterosis among the crosses at the medium-term; the uptrend among traits are wet weight >shell width>shell length> shell height, Chinese cultural population could be recognized as excellent parent, and seasonal variations influence very much on the daily increment and growth rate of each trait of Chlamys farreri and it is only able to survive and could barely grow in winter (6-8 months), but grows fast in temperate season (10-12 months).
基金financially supported by the National Natural Science Foundation of China(31270047)the National High Tech R&D Program(2012AA10A410)+1 种基金the National Basic Research Program of China(2010CB126402)the National Key Technology R&D Program of China(2011BAD45B01 and 2011BAD13B05)
文摘Toll-like receptor(TLR) signaling pathway plays a pivotal role in the innate immune system. Studies on TLR signaling pathway genes in Zhikong scallop(Chlamys farreri) have mainly focused on sequence analysis and expression profiling, no research has been carried out on their localization. The chromosomal position of TLR signaling pathway genes can be valuable for assemblying scallop genome and analysizing gene regulatory networks. In the present study, five key TLR signaling pathway genes(Cf TLR, Cf Myd88, Cf TRAF6, Cf NFκB, and Cf IκB) containing bacterial artificial chromosomes(BACs) were isolated and physically mapped through fluorescence in situ hybridization on five non-homologous chromosome pairs, showing a similar distribution to another five model species. The isolation and mapping of these key immune genes of C. farreri will aid to the research on innate immunity, assignment of interested genes to chromosomes, and integration of physical, linkage and cytogenetic maps of this species.
基金the supports from the National Natural Science Foundation of China(Nos.31130054 and 31472258)the AoShan Talents Program of Qingdao National Laboratory for Marine Science and Technology(No.2015ASTP-ES02)the Fundamental Research Funds for the Central Universities(No.201564009)
文摘This study applied an optimized one-step 2b-RAD library construction strategy and performed simplified genome sequencing of 539 individuals from three continuously selected Chlamys farreri populations. SNP screening was performed using RAD typing software and population genetic parameters for the continuously selected populations from three generations(G1, G2, G3) were determined. The results showed that the optimized one-step 2b-RAD library construction strategy greatly simplified the experimental process, making it suitable for efficiently constructing a large number of libraries. A total of 18450 SNP markers were identified, which evenly distributed throughout the genome. Population genetic analysis of these three generations showed that the mean value of observed heterozygosity was 0.275 ± 0.177, 0.272 ± 0.181 and 0.275 ± 0.166, respectively. Meanwhile, the mean value of expected heterozygosity was 0.275 ± 0.141, 0.274 ± 0.145 and 0.280 ± 0.133, respectively. The Wright's fixation index(F) was 0.04291, 0.04976 and 0.06685, respectively. Markers deviated from Hardy–Weinberg equilibrium accounted for 10.34%, 12.64%, and 23.11%, and the Shannon diversity index was 0.0999 ± 0.0404, 0.0921 ± 0.0388 and 0.0733 ± 0.0308. FIS(also known as the inbreeding coefficient) of the three populations was 0.0256, 0.0323 and 0.0468, respectively. We suggested that the 2b-RAD method is well suited to population genetic studies of aquacultured organisms. Moreover, our results indicated that the continuous selection affected the population genetic structure of the cultured Penglai-Red scallop, but the change was not significant; therefore, population selection should continue.
基金supported by the National High Technology Research and Development Program of China (863 Program) (2012AA10A402)the Natural Science Foundation of Qingdao (11-2-4-1(10)-jch)the Key Natural Science Foundation of Shandong Province (Z2008D02)
文摘DAX1, a member of nuclear receptor superfamily, has a function in the sex determination and gonadal differentiation of several vertebrate species. However, little information about DAX1 of invertebrates is available. Here we cloned a homolog of scallop(Chlamys farreri Jones and Preston 1904) dax1, Cf-dax1, and determined its expression characteristics at m RNA and protein levels. The c DNA sequence of Cf-dax1 was 2093 bp in length, including 1404 bp open reading frame(ORF) encoding 467 amino acids. Unlike those of vertebrates, no conserved LXXLL-related motif was found in the putative DNA binding region of Cf-DAX1. Fluorescence in situ hybridization showed that Cf-dax1 located on the short arm of a pair of subtelocentric chromosomes. Tissue distribution analysis using semi-quantitative RT-PCR revealed that Cf-dax1 expressed widely in adult scallop tissues, with the highest expression level found in adductor muscle, moderate level in mantle, gill and testis, and low level in kidney, ovary and hepatopancreas. The result of quantitative real-time PCR indicated that the expression of Cf-dax1 was significantly higher(P<0.05) in testis than in ovary at the same stage, showing a sex-dimorphic expression pattern. Furthermore, immunohistochemical detection found that Cf-DAX1 mainly located in spermatogonia and spermatocytes of testis and in oogonia and oocytes of ovary, implying that DAX1 may involve in gametogenesis of bivalves.
