Showing papers by "Anand Swaroop published in 1994"
TL;DR: Preliminary studies with cultured human Y79 retinoblastoma cells indicate that expression of PEDF is associated with relatively undifferentiated, proliferating cells rather than their differentiated, slow-growing counterparts, which suggests that PEDf is linked to proliferative events that terminate in final phenotypic determination within specific cell lineages.
82 citations
TL;DR: The amino acid sequence of the SEC13R gene product shows 70% similarity to yeast Sec13p, suggesting thatSEC13R may be the human homolog of SEC13, and the deduced polypeptide sequence contains several beta-transducin like 'WD40' repeats, and is rich in serine and threonine residues.
Abstract: We previously described sequence tags from 58 novel directionally cloned human cDNAs from an enriched retinal pigment epithelial cell line library (Gieser and Swaroop, 1992). The nucleotide sequence of one of the cDNA clones, AA35 (D3S1231E), showed strong homology to the yeast SEC13 gene, required for vesicle biogenesis from endoplasmic reticulum during the transport of proteins. We have designated the human gene SEC13R (SEC13-Related). The amino acid sequence of the SEC13R gene product shows 70% similarity to yeast Sec13p, suggesting that SEC13R may be the human homolog of SEC13. The deduced polypeptide sequence contains several beta-transducin like 'WD40' repeats, and is rich in serine and threonine residues. The 1.4 kb transcript of SEC13R is detected by Northern analysis in many human tissues. However, RT-PCR analysis using two primer sets from different regions of the gene suggests differential expression of alternately spliced transcripts in various tissues. Somatic cell hybrid and in situ hybridization studies localized the SEC13R gene to human chromosome 3p24-p25. A related sequence was mapped to chromosome 18q11.2-q12. SEC13R was physically mapped to a yeast artificial chromosome (YAC) clone spanning the D3S720 marker from the region of the Von Hippel-Lindau disease locus. The mouse Sec13r gene was mapped to the conserved linkage group on chromosome 6 that corresponds to human chromosome 3p24-p25.
32 citations
TL;DR: The results indicate that the peroxidase is not continuously generating H2O2 during scopoletin-mediated NADH oxidation and that both per oxidase and oxidase reactions occur simultaneously competing for an active form of the enzyme.
12 citations
TL;DR: The larger human size mRNA for the human beta 2 Na+,K(+)-ATPase indicates species differences in gene processing.
9 citations
01 Jan 1994
TL;DR: The strategy is now being used for the isolation of cDNAs from large regions of genomic DNA cloned in cosmids or yeast artificial chromosome (YAC) vectors and from microdissected and flow-sorted chromosomal DNA.
Abstract: We are developing a novel strategy for efficient and specific isolation of cDNA clones encoded by a particular genomic region. This sandwich-selection method consists of: 1. Hybridization of single-stranded (ss) circular cDNA molecules to tagged in vitro synthesized RNA from the genomic region; 2. Retention of “genomic RNA” - cDNA hybrids on an avidin-matrix through a biotinylated RNA complementary to the tag; and 3. Elution of specific cDNAs from the avidin-matrix with ribonuclease A. The selected ss cDNAs are directly used for electroporation of competent E. coli cells without requiring any amplification or cloning step. Here, we report the construction of appropriate vectors and results from model experiments. This protocol has been applied for specifically selecting NRL (neural retina leucine zipper) cDNA from a mixture of cDNA clones using a sub-library from the NRL genomic region. The control experiments have allowed us to optimize various steps in the procedure and provided valuable data regarding the yield and specificity of selection process. The strategy is now being used for the isolation of cDNAs from large regions of genomic DNA cloned in cosmids or yeast artificial chromosome (YAC) vectors and from microdissected and flow-sorted chromosomal DNA. We expect that a high degree of specificity and efficiency will permit wider application of this cDNA selection strategy.
2 citations
2 citations