Expression of the dyslexia candidate gene kiaa0319-like in insect cells

The human kiaa0319-like gene is one of the candidate genes for developmental dyslexia, but the exact function of the encoded KIAA0319L (KL) protein is not known. To allow functional analysis a purified, biologically active KL protein is required. The kiaa0319-like gene was expressed in insect cells using the baculovirus expression system. To optimize the expression of the kiaa0319-like gene and to be able to purify the KL protein, several approaches were used. Two different recombinant baculoviruses were made, one with the full length coding sequence of KL and one that lacked the transmembrane domain to facilitate purification.. Versions in which the kiaa0319L sequences were cloned downstream of the honeybee melittin signal sequence were also made. All four constructs contained a C-terminal influenza hemagglutinin (HA)-tag. Sf9 insect cells infected with these recombinant baculoviruses produced the KL protein, as demonstrated by Western blot analysis using either the HA-antibody or KL-specific polyclonal serum

Asthma and atopy are associated with chromosome 17q21 markers in Chinese children

Background: Single‐nucleotide polymorphism (SNP)‐based genome‐wide association study revealed that markers on chromosome 17q21 were linked to childhood asthma but not atopy in Caucasians, with the strongest signal being detected for the SNP rs7216389 in the gene. Such association was unknown in Chinese. This study delineated the allele and genotype frequencies of 10 SNPs at chromosome 17q21, and investigated the relationship between these SNPs and asthma and plasma IgE in southern Chinese children. Methods: Asthmatic children and non‐allergic controls were recruited from pediatric clinics. Their plasma total and aeroallergen‐specific IgE concentrations were measured by immunoassay. Ten SNPs on 17q21 region were genotyped by multiplex SNaPshot™, and their genotype associations with asthma traits analyzed using multivariate regression. Results: 315 patients and 192 controls were enrolled. The allele frequency for C allele of rs7216389 varied significantly from 0.232 in our controls, 0.389 in Han Chinese to 0.536 in Caucasians. Asthma diagnosis was associated with rs11650680 and five other SNPs including rs7216389 (=0.019–0.034), whereas atopy was associated only with rs11650680 (=0.0004). Linear regression revealed the covariates for plasma total IgE to be significant for rs11650680 (=0.008–0.0002). Haplotypic associations were found with atopy and increased plasma total IgE, with the respective odds ratios and 95% confidence intervals for TTTCCGTT haplotype to be 0.21 and 0.09–0.52 (=0.0002) and 0.41 and 0.18–0.90 (=0.025). Conclusion: Childhood asthma and atopy are associated with chromosome 17q21 in Chinese, but such association may involve genes other than in this region