AIM：To investigate the correlation between lumican（LUM）gene and high myopia in a Southern Chinese population.METHODS：The study comprised of 95 high myopia patients with a spherical equivalent≤-6.5 diopters（D）.T...AIM：To investigate the correlation between lumican（LUM）gene and high myopia in a Southern Chinese population.METHODS：The study comprised of 95 high myopia patients with a spherical equivalent≤-6.5 diopters（D）.The control group recruited 95 individuals with a spherical equivalent ranging from-0.5 D to＋0.5 D.Direct sequencing was used to detect the single nucleotide polymorphisms（SNPs）of LUM gene in coding region.Genotype distributions were tested for Hardy-Weinberg disequilibrium.Genotypic and allelic frequencies were analyzed through Chi-square test or Fisher’s exact test.RESULTS：We identified 3 SNPs of the LUM gene：LUM c.32（rs577456426）,LUM c.507（rs17853500）and LUM c.849（rs181915277）.Among the three SNPs,the genotype and allele frequencies of rs17853500 showed a significant difference between patients and control subjects（P〈0.05）.However,there were no significant differences in rs181915277and rs577456426 between the two groups（P〉0.05）.CONCLUSION：LUM c.507 polymorphism may be a risk factor for the pathogenesis of high myopia in the Southern Chinese population.展开更多
AIM： To construct adenovirus vectors of lumican gene by gateway recombinant cloning technology to further understand the role of lumican gene in myopia. METHODS： Gateway recombinant cloning technology was used to co...AIM： To construct adenovirus vectors of lumican gene by gateway recombinant cloning technology to further understand the role of lumican gene in myopia. METHODS： Gateway recombinant cloning technology was used to construct adenovirus vectors. The wild-type （wt） and mutant （mut） forms of the lumican gene were synthesized and amplified by polymerase chain reaction （PCR）. The lumican cDNA fragments were purified and ligated into the adenovirus shuttle vector pDown- multiple cloning site （MCS）-/internal ribozyme entry site （IRES）/enhanced green fluorescent protein （EGFP）. Then the desired DNA fragments were integrated into the destination vector pAV.Desld yielding the final expression constructs pAV.Exld-CMV〉wt-lumican/IRES/ EGFP and pAV.Exld-cytomegalovirus （CMV） 〉mutlumican/IRES/EGFP, respectively.RESULTS： The adenovirus plasmids pAV.Exld-CMV〉 wt-lumican/IRES/EGFP and pAV.Exld-CMV 〉mutlumican/IRESlEGFP were successfully constructed by gateway recombinant cloning technology. Positive clones identified by PCR and sequencing were selected and packaged into recombinant adenovirus in HEK293 cells. CONCLUSION： We construct adenovirus vectors containing the lumican gene by gateway recombinant cloning technology, which provides a basis for investigating the role of lumicangene in the pathogenesis of high myopia.展开更多
基金Supported by Natural Science Foundation of Guangdong Province,China(No.2015A030310158No.2014A030313359)+1 种基金the Science and Technology Planning Project of Guangdong Province,China(No.2015B020226003)the Scientific and Cultivation Foundation of the First Affiliated Hospital of Jinan University(No.2015201)
文摘AIM：To investigate the correlation between lumican（LUM）gene and high myopia in a Southern Chinese population.METHODS：The study comprised of 95 high myopia patients with a spherical equivalent≤-6.5 diopters（D）.The control group recruited 95 individuals with a spherical equivalent ranging from-0.5 D to＋0.5 D.Direct sequencing was used to detect the single nucleotide polymorphisms（SNPs）of LUM gene in coding region.Genotype distributions were tested for Hardy-Weinberg disequilibrium.Genotypic and allelic frequencies were analyzed through Chi-square test or Fisher’s exact test.RESULTS：We identified 3 SNPs of the LUM gene：LUM c.32（rs577456426）,LUM c.507（rs17853500）and LUM c.849（rs181915277）.Among the three SNPs,the genotype and allele frequencies of rs17853500 showed a significant difference between patients and control subjects（P〈0.05）.However,there were no significant differences in rs181915277and rs577456426 between the two groups（P〉0.05）.CONCLUSION：LUM c.507 polymorphism may be a risk factor for the pathogenesis of high myopia in the Southern Chinese population.
基金Supported by the Natural Science Foundation of Guangdong Province(No.2015A030310158No.2014A030313359)+1 种基金the Fundamental Research Funds for the Central Universities(No.21611446)the Scientific and Cultivation Foundation of the First Affiliated Hospital of Jinan University(No.2015201)
文摘AIM： To construct adenovirus vectors of lumican gene by gateway recombinant cloning technology to further understand the role of lumican gene in myopia. METHODS： Gateway recombinant cloning technology was used to construct adenovirus vectors. The wild-type （wt） and mutant （mut） forms of the lumican gene were synthesized and amplified by polymerase chain reaction （PCR）. The lumican cDNA fragments were purified and ligated into the adenovirus shuttle vector pDown- multiple cloning site （MCS）-/internal ribozyme entry site （IRES）/enhanced green fluorescent protein （EGFP）. Then the desired DNA fragments were integrated into the destination vector pAV.Desld yielding the final expression constructs pAV.Exld-CMV〉wt-lumican/IRES/ EGFP and pAV.Exld-cytomegalovirus （CMV） 〉mutlumican/IRES/EGFP, respectively.RESULTS： The adenovirus plasmids pAV.Exld-CMV〉 wt-lumican/IRES/EGFP and pAV.Exld-CMV 〉mutlumican/IRESlEGFP were successfully constructed by gateway recombinant cloning technology. Positive clones identified by PCR and sequencing were selected and packaged into recombinant adenovirus in HEK293 cells. CONCLUSION： We construct adenovirus vectors containing the lumican gene by gateway recombinant cloning technology, which provides a basis for investigating the role of lumicangene in the pathogenesis of high myopia.
