ABSTRACT
OBJECTIVE@#To identify the blood group of a patient with DEL phenotype combined with positive direct anti-human globulin test and to analyze the pedigree.@*METHODS@#Routine serological reagents were used for serological analysis of RhD blood group in the pedigree members. Exons and flanking sequences of RHD gene were amplified, sequenced and analyzed for heterozygosity. The familial genetic state of DEL phenotype was further analyze in the family members.@*RESULTS@#The DAT was strongly positive in the proband. The 1227G>A allele (RHD*DEL1) was present in the exon 9 of RHD gene, and the mother was the carrier of RHD*DEL1. The proband was identified as RHD+/RHD-, suggesting the CDe/Cde haplotype.@*CONCLUSION@#The proband is DEL phenotype (RHD*DEL1).
Subject(s)
Humans , Alleles , Exons , Genotype , Pedigree , Phenotype , Rh-Hr Blood-Group SystemABSTRACT
OBJECTIVE@#To investigate the phenotype and genotype of the weak D blood group in one case of Chinese Han people.@*METHODS@#Phenotype of blood sample was identified with serologic tests; PCR-SBT was applied for the analysis of genotype and RhD zygosity.@*RESULTS@#Both saline and gel card tests demonstrated this case to be dCcee, which was contrary to glass bead card result. Some of the RBC D epitopes were negative.c.1022T>A allele was identified with PCR-SBT and the zygosity analysis showed this case to be D/d.@*CONCLUSION@#RHD*1022 A is more suitable to be categorized as weak partial D other than weak D in a Chinese Han people.
Subject(s)
Humans , Alleles , Asian People , Exons , Genotype , Phenotype , Rh-Hr Blood-Group SystemABSTRACT
<p><b>OBJECTIVE</b>To clone the circular RNA hsa_circ_0000254 and construct its lentiviral over-expression vector.</p><p><b>METHODS</b>The sequence of hsa_circ_0000254 (a total of 524 bp long) was synthesized and cloned by using pGH vector. The vector was cut by EcoR I and BamH I, and artificial hsa_circ_0000254 was obtained, then inserted in pLCDH-ciR to construct the recombinant expression vector pLCDH-circ254(C254), which was confirmed by DNA sequencing. The lentiviral expression vectors pLCDH-circ254(C254) and NC(pLCDH-ciR) were cotransfected into 293T cells by lipofectamine 2000(lipo2000). After transfection for 40 hours, the cells were collected and verified by PCR and sequencing.</p><p><b>RESULTS</b>Restriction analysis and DNA sequencing demonstrated that the lentiviral vector pLCDH-circ254(C254) was constructed successfully, the expression efficiency increased 10000 times after transfection of cells.</p><p><b>CONCLUSION</b>The successful construction of the lentiviral expression vector pLCDH-circ254(C254) results in the production of high-titer lentivirus, so as to facilitate further study of the molecular functions of hsa_circ_0000254.</p>