By homologous expressed sequence tag (EST) searching, one EST (GenBank: W29095) was obtained, which shows 75% identity in 435 bp overlap with the coding sequence of mouse Cacng2 gene. A 1 545 bp cDNA fragment was obta...By homologous expressed sequence tag (EST) searching, one EST (GenBank: W29095) was obtained, which shows 75% identity in 435 bp overlap with the coding sequence of mouse Cacng2 gene. A 1 545 bp cDNA fragment was obtained from the nested polymerase chain reaction (PCR) and rapid applification of cDNA end (RACE) reaction in the human brain prefrontal cortex cDNA library and the human brain Ready cDNA with the primers designed on W29095. The fragment contained a 948-bp open reading frame (ORF) encoding 315 amino acids, and was named CACNG3. As it was identical to a BAC clone (GenBank: AC004125) from chromosome 16p12-p13.1, the CACNG3 gene was mapped to human chromosome 16p12-p13.1, and the coding region was composed of 4 exons. Reverse transcription PCR (RT-PCR) analysis showed that the CACNG3 gene expressed in human adult brain and fetal brain. Single strand comformation polymorphism (SSCP) analysis was performed in 3 pedigrees with autosomal recessive retinitis pigmentosa, 8 pedigrees with autosomal展开更多
5 μg of total RNAs from normal nasopharynx and nasopharyngeal carcinoma tissue have been labeled with α-32P-dCTP during reverse transcription. The synthesized cDNA probes have been hybridized to high-density cDNA mi...5 μg of total RNAs from normal nasopharynx and nasopharyngeal carcinoma tissue have been labeled with α-32P-dCTP during reverse transcription. The synthesized cDNA probes have been hybridized to high-density cDNA microarray containing 5184 genes or expression sequence tags (ESTs). Then image analysis software has been applied to comparing their expression profiles. Results show that 187 ESTs were of density value above 200 in nasopharyngeal carcinoma tissue while there were 307 such ESTs in normal nasopharynx tissue; 38 ESTs were strongly expressed in nasopharynx, but weakly expressed in nasopharyngeal carcinoma; 48 ESTs were strongly expressed in nasopharyngeal carcinoma, but weakly expressed in normal nasopharynx. These results suggest that there may exist some new differentially expressed genes involved in nasopharyngeal carcinoma development. Furthermore, the results strongly indicate that high-density cDNA microarray is a powerful and efficient tool for large-scale screening differentially展开更多
One EST N27741 with high expression in normal adult nasopharynx tissues but low expression in adult poorly differentiated squamous nasopharyngeal carcinoma has been selected out by the high-density cDNA array expressi...One EST N27741 with high expression in normal adult nasopharynx tissues but low expression in adult poorly differentiated squamous nasopharyngeal carcinoma has been selected out by the high-density cDNA array expression profiling technique. The differential expression has been confirmed by RT-PCR. One novel gene of 1096 bp has been cloned based on this EST. Bioinformatics analysis found that the new gene sequence contains a whole reading frame encoding 256 amino acids. There is a stop codon TAA in front of the 5’ end start codon, and a tailing signal AATAAA and poly A tail at the 3’ end. There is no homologous known gene found after searching by blasting this sequence to non-redundancy nucleotide database. Therefore it is considered a novel gene related to nasopharyngeal carcinoma.展开更多
Using a synthetic oligonucleotide (dA-dC)<sub>15</sub> as a probe to screen a probe pool made by microdissected human chromosome band 17q11-q12, a DNA fragment containing (CA)<sub>16</sub> ...Using a synthetic oligonucleotide (dA-dC)<sub>15</sub> as a probe to screen a probe pool made by microdissected human chromosome band 17q11-q12, a DNA fragment containing (CA)<sub>16</sub> was isolated, which is a novel short tandem repeat (STR) determined by searching in the GenBank and GDB. It has 8 allelic types with a PIC value of 0.61. The novel STR is conformed with Mendelian segregation according to the linkage analysis of a three-generation neurofibromatosis type Ⅰ (NF1) pedigree. The STR was localized at chromosome band 17q11-q12 in the vicinity of NF1 gene by FISH analysis. The accession number of the STR in GenBank and GDB is G32112 and D17S2204 respectively.展开更多
文摘By homologous expressed sequence tag (EST) searching, one EST (GenBank: W29095) was obtained, which shows 75% identity in 435 bp overlap with the coding sequence of mouse Cacng2 gene. A 1 545 bp cDNA fragment was obtained from the nested polymerase chain reaction (PCR) and rapid applification of cDNA end (RACE) reaction in the human brain prefrontal cortex cDNA library and the human brain Ready cDNA with the primers designed on W29095. The fragment contained a 948-bp open reading frame (ORF) encoding 315 amino acids, and was named CACNG3. As it was identical to a BAC clone (GenBank: AC004125) from chromosome 16p12-p13.1, the CACNG3 gene was mapped to human chromosome 16p12-p13.1, and the coding region was composed of 4 exons. Reverse transcription PCR (RT-PCR) analysis showed that the CACNG3 gene expressed in human adult brain and fetal brain. Single strand comformation polymorphism (SSCP) analysis was performed in 3 pedigrees with autosomal recessive retinitis pigmentosa, 8 pedigrees with autosomal
文摘5 μg of total RNAs from normal nasopharynx and nasopharyngeal carcinoma tissue have been labeled with α-32P-dCTP during reverse transcription. The synthesized cDNA probes have been hybridized to high-density cDNA microarray containing 5184 genes or expression sequence tags (ESTs). Then image analysis software has been applied to comparing their expression profiles. Results show that 187 ESTs were of density value above 200 in nasopharyngeal carcinoma tissue while there were 307 such ESTs in normal nasopharynx tissue; 38 ESTs were strongly expressed in nasopharynx, but weakly expressed in nasopharyngeal carcinoma; 48 ESTs were strongly expressed in nasopharyngeal carcinoma, but weakly expressed in normal nasopharynx. These results suggest that there may exist some new differentially expressed genes involved in nasopharyngeal carcinoma development. Furthermore, the results strongly indicate that high-density cDNA microarray is a powerful and efficient tool for large-scale screening differentially
文摘One EST N27741 with high expression in normal adult nasopharynx tissues but low expression in adult poorly differentiated squamous nasopharyngeal carcinoma has been selected out by the high-density cDNA array expression profiling technique. The differential expression has been confirmed by RT-PCR. One novel gene of 1096 bp has been cloned based on this EST. Bioinformatics analysis found that the new gene sequence contains a whole reading frame encoding 256 amino acids. There is a stop codon TAA in front of the 5’ end start codon, and a tailing signal AATAAA and poly A tail at the 3’ end. There is no homologous known gene found after searching by blasting this sequence to non-redundancy nucleotide database. Therefore it is considered a novel gene related to nasopharyngeal carcinoma.
文摘Using a synthetic oligonucleotide (dA-dC)<sub>15</sub> as a probe to screen a probe pool made by microdissected human chromosome band 17q11-q12, a DNA fragment containing (CA)<sub>16</sub> was isolated, which is a novel short tandem repeat (STR) determined by searching in the GenBank and GDB. It has 8 allelic types with a PIC value of 0.61. The novel STR is conformed with Mendelian segregation according to the linkage analysis of a three-generation neurofibromatosis type Ⅰ (NF1) pedigree. The STR was localized at chromosome band 17q11-q12 in the vicinity of NF1 gene by FISH analysis. The accession number of the STR in GenBank and GDB is G32112 and D17S2204 respectively.