Molecular Characterization, Developmental Expression and Immunolocalization of Clathrin Heavy Chain in the Ovary of the American Cockroach, Periplaneta Americana During Oogenesis

Clathrin is the principal protein involved in receptor mediate endocytosis and the main component of the coated vesicles. It is composed of three identical clathrin heavy chains (CHC), each with an attached light chain. We characterized the deduced amino acid sequence of the partial cDNA clone of the American cockroach, Periplaneta americana (Pam) CHC. The analysis showed that this sequence is represented as multiple alpha helical repeats occurred in the arm region of the CHC and displayed a high level of identity and similarity to mosquitoes and Drosophila melanogaster CHCs. This is the first report on CHC from a hemimetabolous insect. The amplified CHC probe could hybridize two CHC transcripts in the current preparations, 6.3 kb and 7.3 kb. The Northern blot analysis confirmed that a 6.3 kb transcript is specifically expressed in ovarian tissues at high levels throughout the ovarian development, especially in previtellogenic ovaries (Days 1-4) but dropped during the vitellogenic period (days 5-7) and ultimately no transcript was detected in fully vitellogenic ovaries (days 9-13). Immunoblot analysis detected an ovary specific CHC protein of ~175 kDa that was present in previtellogenic ovaries on the day of female emergence and after initiation of vitellogenesis and onset of Vg uptake. Immunocytochemistry localized CHC protein to germ-line derived cells, oocytes, and revealed that CHC translation begins very early during oocyte differentiation in the germarium. The present work suggested a possible role for clathrin in the early fluid phase endocytosis (pinocytosis) in addition to its role in receptor-mediated endocytosis

Molecular characterization and DNA fingerprinting of some local eggplant genotypes and its wild relatives

Collection and characterization of local genotypes and landraces are prerequisite for any crop improvement program. Molecular diversity and DNA profiling shown exact genetic blue print of any crop. Hence, the experiment was design to establish the molecular diversity and polymorphism among some local eggplant genotypes and its wild relatives for future breeding program. The experiment was carried out at the Biotechnology Laboratory, Sher-e-Bangla Agricultural University, Dhaka, Bangladesh, with twenty-five local and two wild relatives (Solanum sisymbriifolium and S. villosum) of eggplant to study molecular diversity and DNA fingerprinting at those genotypes. Five well-known SSR primers (EPSSR82, smSSR01, EM114, EM120 and smSSR04) were used for the molecular characterization of the genotypes. Quality DNA was isolated with 27 genotypes and PCR amplification was carried out with these primer. The amplified DNA fragment was visualized by 2% agarose gel and data were analyzed by POWERMAKER (version 3.25) and NTSYS-PC (version 2.2). Some total at 10 different alleles were generated with a range of 1 to 3 alleles per locus and an average of 2.0 alleles. The highest number (2) of polymorphic bands was observed in the primers EPSSR82 and smSSR01. The Polymorphism Information Content (PIC) of SSR markers ranged from 0.37 to 0.67 with an average value of PIC = 0.54. Gene diversity ranges from 0.49 (smSSR01) to 0.72 (EPSSR82), with an average value of 0.61. UPGMA method separated the of 27 genotypes into two major clusters (I and II). From the clusters, wild species Solanum villosum belonged to the sub-cluster (IIb), that revealed its distinct variation from the others. On the other hand, wild species Solanum sisymbriifolium showed a close relatedness by forming the same cluster together with thirteen local eggplant genotypes. Molecular diversity and DNA profiling was identified among 25 local eggplant germplasm and its wild relatives. The finding of the experiment could be used for selection of diverse parent for eggplant improvement

Molecular Characterization and Study of Genetic Relationships among local Cultivars of the Moroccan fig (Ficus carica L.) using Microsatellite and ISSR Markers

Molecular characterization of Moroccan local fig (Ficus carica L.) germplasm was performed on the cultivars present in a collection of the National School of Agriculture of Meknes. A total of 22 fig samples were analysed using 7 ISSR primers and 9 loci S.S.R. A total of 54 I.S.S.R. polymorphic bands with an average of 8 per primers and 42 S.S.R. alleles with means 5 alleles per locus were revealed by these analyses. The ISSR markers allowed distinguishing 22 molecular profiles and S.S.R. loci differentiated between 21 different profiles. Pairwise Comparing, 87% of cultivars pairs were differentiated by 7 to 24 alleles and 89% by 9 to 29 ISSR bands. The statistical analysis and genetic distances have shown a wide molecular diversity in the collection, where the average observed heterozygosity was 0.42. The average similarity between cultivars is 70% using SSR markers and 71.6 for ISSR markers. The same SSR profile was obtained for Nabout1 and Nabout2 with 0 allele difference. Small differences of 1 to 6 alleles were obtained among cultivars which have the same names, which presumably corresponds to somaclonal variations obtained through intense vegetative propagation over long periods, while the differences over 7 alleles suggests the problems of homonyms

Molecular Characterization of Nepali Potato Cultivars Using Randomly Amplified Polymorphic DNA (Rapd) Markers

Randomly amplified polymorphic DNA (RAPD) was used to study the genetic diversity of four local cultivars of potato. Amplification with ten arbitrary decamer primers produced 29 different marker bands of which 69.0% were polymorphic. The size range of the amplified DNAs ranged between 370 bp and 2500 bp. On average, 17.5 alleles per genotype were amplified using the RAPD primers. With the selected primers sufficient polymorphism could be detected to allow identification of individual genotypes. A dendrogram displaying the relative genetic similarities between the genotypes showed a range of 55.2-69.0% similarity

Molecular characterization and systemic induction of single-chain ribosome-inactivating proteins (RIPs) in sugar beet (Beta vulgaris) leaves

Producción CientíficaSugar beet (Beta vulgaris L.) leaves contain virusinducible type 1 (single chain) ribosome-inactivating proteins that have been named beetins. The structural and functional characterization, the cellular location, and the potential role of beetins as antiviral agents are reported here. Beetins are formed of a single polypeptide chain with a varying degree of glycosylation and strongly inhibited in vitro protein synthesis in rabbit reticulocyte lysates (IC5051.15 ng ml21 ) and a Vicia sativa L. cell-free system (IC50568 ng ml21 ) through the single depurination of the large rRNA. Beetins trigger the multidepurination of tobacco mosaic virus (TMV) genomic RNA which underwent extensive degradation upon treatment with acid aniline. Beetins are extracellular proteins that were recovered from the apoplastic fluid. Induction of sugar beet RIPs with either H2O2 or artichoke mottled crinkle virus (AMCV) was observed in leaves distant from the site of application of such elicitors. The external application of purified beetin to sugar leaves prevented infection by AMCV which supports the preliminary hypothesis that beetins could be involved in plant systemic acquired resistance subjected to induction by phytopathogens.Comisión Interministerial de Ciencia y Tecnología (grant BIO98-0727)Junta de Castilla y León - FIS (grant PI030258

A note on HwpH^p_w-boundedness of Riesz transforms and θ\theta-Calder\'on-Zygmund operators through molecular characterization

Let $0 < p \leq 1$ and $w$ in the Muckenhoupt class $A_1$. Recently, by using the weighted atomic decomposition and molecular characterization; Lee, Lin and Yang \cite{LLY} (J. Math. Anal. Appl. 301 (2005), 394--400) established that the Riesz transforms $R_j, j=1, 2,...,n$, are bounded on $H^p_w(\mathbb R^n)$. In this note we extend this to the general case of weight $w$ in the Muckenhoupt class $A_\infty$ through molecular characterization. One difficulty, which has not been taken care in \cite{LLY}, consists in passing from atoms to all functions in $H^p_w(\mathbb R^n)$. Furthermore, the $H^p_w$-boundedness of $\theta$-Calder\'on-Zygmund operators are also given through molecular characterization and atomic decomposition.Comment: to appear in Anal. Theory. Appl. 27 (2011), no. 3, 251-26

Phenotypic and molecular assessment of seven patients with 6p25 deletion syndrome: Relevance to ocular dysgenesis and hearing impairment

BACKGROUND: Thirty-nine patients have been described with deletions involving chromosome 6p25. However, relatively few of these deletions have had molecular characterization. Common phenotypes of 6p25 deletion syndrome patients include hydrocephalus, hearing loss, and ocular, craniofacial, skeletal, cardiac, and renal malformations. Molecular characterization of deletions can identify genes that are responsible for these phenotypes. METHODS: We report the clinical phenotype of seven patients with terminal deletions of chromosome 6p25 and compare them to previously reported patients. Molecular characterization of the deletions was performed using polymorphic marker analysis to determine the extents of the deletions in these seven 6p25 deletion syndrome patients. RESULTS: Our results, and previous data, show that ocular dysgenesis and hearing impairment are the two most highly penetrant phenotypes of the 6p25 deletion syndrome. While deletion of the forkhead box C1 gene (FOXC1) probably underlies the ocular dysgenesis, no gene in this region is known to be involved in hearing impairment. CONCLUSIONS: Ocular dysgenesis and hearing impairment are the two most common phenotypes of 6p25 deletion syndrome. We conclude that a locus for dominant hearing loss is present at 6p25 and that this locus is restricted to a region distal to D6S1617. Molecular characterization of more 6p25 deletion patients will aid in refinement of this locus and the identification of a gene involved in dominant hearing loss

Molecular Characterization of Three Cultivars of Tomato (Lycopersicon Esculentum L.) in South-West Nigeria Using SSR Markers

Molecular characterisation of local tomato cultivars – Ibadan Local (IbL), Ife and JM94/46 (JM) were assessed using simple sequence repeat (SSR) markers. Out of ten SSR primer pairs used, three primer pairs were able to differentiate amplified genomic DNA of the cultivars. Unweighted Pair Group Method Using Arithmetic Average (UPGMA) cluster analysis of the data showed a close relationship between IbL and Ife with a genetic distance (GD) of 0.067; Ife and JM had GD of 0.2 and JM and Ife had GD of 0.25

Molecular characterization of Cryptosporidium and Giardia in farmers and their ruminant livestock from the Coastal Savannah zone of Ghana

Cryptosporidium and Giardia are major causes of diarrhoea in developing countries including Ghana, however, nothing is known about the species and subtypes of Cryptosporidium and Giardia in farmers and their ruminant livestock in this country. A total of 925 faecal samples from humans (n = 95), cattle (n = 328), sheep (n = 217) and goats (n = 285), were screened for Cryptosporidium and Giardia by quantitative PCR (qPCR) at the 18S rRNA and glutamate dehydrogenase (gdh) loci respectively. Cryptosporidium positives were typed by sequence analysis of 18S and 60 kDa glycoprotein (gp60) loci amplicons. Giardia positives were typed at the triose phosphate isomerase (tpi), beta-giardin (bg) and gdh loci. The prevalence of Cryptosporidium and Giardia by qPCR was 8.4% and 10.5% in humans, 26.5% and 8.5% in cattle, 34.1% and 12.9% in sheep, and 33.3% and 12.3% in goat faecal samples, respectively. G. duodenalis assemblages A and B were detected in humans and assemblage E was detected in livestock. Cryptosporidium parvum was the only species identified in humans; C. andersoni, C. bovis, C. ryanae and C. ubiquitum were identified in cattle; C. xiaoi, C. ubiquitum and C. bovis in sheep; and C. xiaoi, C. baileyi and C. parvum in goats. This is the first molecular study of Cryptosporidium and Giardia in livestock in Ghana. The identification of zoonotic species and the identification of C. parvum subtype IIcA5G3q in livestock, which has previously been identified in children in Ghana, suggests potential zoonotic transmission. Further studies on larger numbers of human and animal samples, and on younger livestock are required to better understand the epidemiology and transmission of Cryptosporidium and Giardia in Ghana