Polymorphism of follicle stimulating hormone beta sub-unit (FSH-P) gene as a molecular marker for reproductive status in Peranakan Ongole x Bali crossbred (POBA) cattle

Abstract POBA cattle is a crossbred cattle from Peranakan Ongole (PO) and Bali cattle, which is developed by the Beef Cattle Research Institute (BCRI), Ministry of Agriculture of the Republic of Indonesia. This study aimed to identify the genetic and phenotypic characteristics of POBA cattle raised in the BCRI. Genetic diversity of FSH-β gene was also determined in order to identify a possible marker for reproductive status in POBA cattle. Rambon cattle collected from Banyuwangi regency were used as comparison. A 313 bp fragment of the FSH-β gene was amplified using the polymerase chain reaction (PCR). The PCR products were sequenced and aligned to detect polymorphism. As results, a polymorphism (SNP g.2583C/T) in the FSH-β gene, which produced three genotypes (TT, CC, and CT) was detected. The frequency of each allele was 0.13 (allele C) and 0.87 (allele T). However, the FSH-β gene polymorphism did not significantly affect sperm quality, body weight, body size, and cervical condition of POBA cattle.

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A Comparative Study of 5S rDNA Non-Transcribed Spacers in Elaeagnaceae Species

Plants ◽

10.3390/plants10010004 ◽

2020 ◽

Vol 10 (1) ◽

pp. 4

Author(s):

Oleg S. Alexandrov ◽

Olga V. Razumova ◽

Gennady I. Karlov

Keyword(s):

Polymerase Chain Reaction ◽

Molecular Markers ◽

Dna Markers ◽

5S Rdna ◽

Chain Reaction ◽

Closely Related Species ◽

Pcr Products ◽

Polymerase Chain ◽

Species Specific ◽

Shepherdia Canadensis

5S rDNA is organized as a cluster of tandemly repeated monomers that consist of the conservative 120 bp coding part and non-transcribed spacers (NTSs) with different lengths and sequences among different species. The polymorphism in the 5S rDNA NTSs of closely related species is interesting for phylogenetic and evolutional investigations, as well as for the development of molecular markers. In this study, the 5S rDNA NTSs were amplified with universal 5S1/5S2 primers in some species of the Elaeagnaceae Adans. family. The polymerase chain reaction (PCR) products of five Elaeagnus species had similar lengths near 310 bp and were different from Shepherdia canadensis (L.) Nutt. and Sh. argentea (Pusch.) Nutt. samples (260 bp and 215 bp, respectively). The PCR products were cloned and sequenced. An analysis of the sequences revealed that intraspecific levels of NTS identity are high (approximately 95–96%) and similar in the Elaeagnus L. species. In Sh. argentea, this level was slightly lower due to the differences in the poly-T region. Moreover, the intergeneric and intervarietal NTS identity levels were studied and compared. Significant differences between species (except E. multiflora Thunb. and E. umbellata Thunb.) and genera were found. Herein, a range of the NTS features is discussed. This study is another step in the investigation of the molecular evolution of Elaeagnaceae and may be useful for the development of species-specific DNA markers in this family.

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Automated closed-vessel system for in vitro diagnostics based on polymerase chain reaction

Clinical Chemistry ◽

10.1093/clinchem/39.9.1927 ◽

1993 ◽

Vol 39 (9) ◽

pp. 1927-1933 ◽

Cited By ~ 39

Author(s):

J B Findlay ◽

S M Atwood ◽

L Bergmeyer ◽

J Chemelli ◽

K Christy ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Pcr Amplification ◽

Automated System ◽

Closed Vessel ◽

Chain Reaction ◽

Panel Testing ◽

Pcr Product ◽

Pcr Products ◽

Polymerase Chain

Abstract An automated system for polymerase chain reaction (PCR) amplification and detection combats false-positive results caused by "PCR product carryover." The system uses a single vessel for both PCR amplification and the subsequent detection of PCR products, eliminating the need to handle PCR products in an open environment and risk product carryover. The sample and PCR reagents are introduced into one compartment within the vessel, and amplification occurs as they are thermally cycled. Other compartments contain the reagents for detection of PCR products. Pressure from a roller provides for sequential delivery of the contents of the compartments to a detection area. The PCR products are biotinylated at their 5' ends during amplification through the use of biotinylated primers. After delivery to the detection area, they are specifically captured by hybridization with immobilized oligonucleotide probes. Subsequent reaction with streptavidin-horseradish peroxidase conjugate forms a complex that catalyzes dye formation from dye precursor. Wash steps minimize nonspecific background. This format is amenable to multiplexing, permitting internal controls, speciation of bacteria, typing of viruses, and panel testing. An HIV assay performed with this system demonstrated 100% sensitivity and 95% specificity for 64 patients' samples relative to a conventional PCR assay based on 32P solution hybridization. Similarly, an automated closed-vessel assay of cytomegalovirus exhibited 97.5% sensitivity and 100% specificity.

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Fluorometric Polymerase Chain Reaction (PCR) Enzyme-Linked Immunosorbent Assay for Quantification of Immuno-PCR Products in Microplates

Analytical Biochemistry ◽

10.1006/abio.1996.9989 ◽

1997 ◽

Vol 246 (1) ◽

pp. 140-145 ◽

Cited By ~ 61

Author(s):

Christof M. Niemeyer ◽

Michael Adler ◽

Dietmar Blohm

Keyword(s):

Polymerase Chain Reaction ◽

Immunosorbent Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Chain Reaction ◽

Pcr Products ◽

Polymerase Chain

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PCR-induced (ligase-free) subcloning: a rapid reliable method to subclone polymerase chain reaction (PCR) products

Nucleic Acids Research ◽

10.1093/nar/18.7.1920 ◽

1990 ◽

Vol 18 (7) ◽

pp. 1920-1920 ◽

Cited By ~ 36

Author(s):

Alan R. Shuldiner ◽

Laurie A. Scott ◽

Jesse Roth

Keyword(s):

Polymerase Chain Reaction ◽

Reliable Method ◽

Chain Reaction ◽

Pcr Products ◽

Polymerase Chain

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Detection of the chromosomal translocation t(11;14) by polymerase chain reaction in mantle cell lymphomas

Blood ◽

10.1182/blood.v83.7.1871.1871 ◽

1994 ◽

Vol 83 (7) ◽

pp. 1871-1875 ◽

Cited By ~ 150

Author(s):

R Rimokh ◽

F Berger ◽

G Delsol ◽

I Digonnet ◽

JP Rouault ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Chromosomal Translocation ◽

Nucleotide Sequencing ◽

Chromosome 11 ◽

Chromosome 14 ◽

Chain Reaction ◽

Mantle Cell ◽

Pcr Products ◽

Polymerase Chain ◽

Ig Heavy Chain

Abstract The t(11;14)(q13;q32) and its molecular counterpart, BCL1 rearrangement, are consistent features of mantle cell lymphoma (MCL). Rearrangement is thought to deregulate the nearby CCND1 (BCL1/PRAD1) proto-oncogene, a member of the cyclin G1 gene family, and thereby to contribute to tumorigenesis. We and others have previously shown that the BCL1 locus is rearranged in 55% to 60% of MCL patients and that, on chromosome 11, more than 80% of the breakpoints are localized within a 1-kbp DNA segment known as the major translocation cluster (MTC). We have determined the nucleotide sequence for a portion of the MTC region, and constructed chromosome 11-specific oligonucleotides that were in conjunction with a consensus immunoglobulin (Ig) heavy chain joining region (JH) primer used to perform the polymerase chain reaction (PCR) to amplify t(11;14) chromosomal junctional sequences in DNA from 16 MCL patients with breakpoints in the MTC region. 15 of the 16 breakpoints that occurred at the MTC region were amenable to PCR detection. The sizes of the amplified bands, the existence or not of a Sac I site in the PCR products, and nucleotide sequencing of the amplified DNA from four patients showed that the breakpoints share a remarkable tendency to tightly cluster within 300 bp on chromosome 11, some of them occurring at the same nucleotide. On chromosome 14, the breakpoints were localized within the Ig JH. Our findings indicate that a BCL1 rearrangement can be detected using this approach in roughly one half of the MCL patients. This has implications for both the diagnosis and the clinical management of MCL.

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Development of an EST-SSR marker in Panax ginseng

Chinese Journal of Agricultural Biotechnology ◽

10.1017/s1479236208002167 ◽

2008 ◽

Vol 5 (2) ◽

pp. 175-181 ◽

Cited By ~ 6

Author(s):

Yang Cheng-Jun ◽

Wang Jun ◽

Mu Li-Qiang ◽

Li Shao-Chen ◽

Liu Guan-Jun ◽

...

Keyword(s):

Panax Ginseng ◽

Expressed Sequence Tags ◽

Genomic Dna ◽

Ssr Marker ◽

Panax Quinquefolius ◽

Chain Reaction ◽

Acanthopanax Senticosus ◽

Pcr Products ◽

Polymerase Chain ◽

Expressed Sequence

AbstractA total of 791 microsatellites (SSRs) were isolated from 7055 Panax ginseng expressed sequence tags (ESTs). According to primer design criteria, 68 primer pairs for EST-SSR were designed. Under an appropriate polymerase chain reaction (PCR) system, all EST-SSR primer pairs were screened against genomic DNA of Ji'anchangbo and Fusong'ermaya from Panax ginseng, and 43 EST-SSR primer pairs out of the above 68 resulted in PCR products. Then, all 43 pairs were detected in nine P. ginseng, two Panax quinquefolius and two Acanthopanax senticosus cultivars for polymorphisms, and 26 pairs (60.47%) were found to be polymorphic, accounting for 38.23% of the total number of designed primer pairs. These results demonstrate the possibility of developing EST-SSR markers using P. ginseng ESTs.

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Acrosome reaction-inducing substance triggers two different pathways of sperm intracellular signaling in newt fertilization

The International Journal of Developmental Biology ◽

10.1387/ijdb.190092aw ◽

2019 ◽

Vol 63 (11-12) ◽

pp. 589-595

Author(s):

Shinnosuke Kon ◽

Akio Takaku ◽

Fubito Toyama ◽

Eriko Takayama-Watanabe ◽

Akihiko Watanabe

Keyword(s):

Intracellular Signaling ◽

Acrosome Reaction ◽

Sperm Quality ◽

Adenylyl Cyclases ◽

Chain Reaction ◽

Gated Channel ◽

Polymerase Chain ◽

Intracellular Pathways ◽

Intracellular Mediators ◽

Kinase A

The acrosome reaction is induced in the sperm of Cynops pyrrhogaster immediately in response to a ligand protein called acrosome reaction-inducing substance (ARIS) in the egg jelly at fertilization, whereas a spontaneous acrosome reaction occurs time-dependently in correlation with the decline of sperm quality for fertilization. The ARIS-induced acrosome reaction was recently found to be mediated by TRPV4 in association with the NMDA type glutamate receptor, although the intracellular mediators for the acrosome reaction are largely unclear. In the present study, spontaneous acrosome reaction was significantly inhibited by Ni2+, RN1734, and diltiazem, which blocks Cav3.2, TRPV4 or TRPM8, and the cyclic nucleotide-gated channel, respectively. In contrast, expression of Ca2+-activated transmembrane and soluble adenylyl cyclases was detected in the sperm of C. pyrrhogaster by reverse transcription-polymerase chain reaction. Activator of transmembrane or soluble adenylyl cyclases (forskolin or HCO3-) independently promoted spontaneous acrosome reaction, while an inhibitor of each enzyme (MD12330A or KH7) inhibited it only in the sperm with high potential for spontaneous acrosome reaction. An inhibitor of protein kinase A (H89) inhibited spontaneous acrosome reaction in a manner independent of sperm potential for spontaneous acrosome reaction. Surprisingly, KH7 significantly inhibited ARIS-induced acrosome reaction, but its effect was seen in a small percentage of sperm. H89 had no effect on ARIS-induced acrosome reaction. These results suggest that C. pyrrhogaster sperm possess multiple intracellular pathways for acrosome reaction, involving Ca2+ permeable channels, adenylyl cyclases and PKA, and that two pathways having distinct dependencies on adenylyl cyclases may contribute to ARIS-induced acrosome reaction at fertilization.

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Detection of Subclinical Systemic Disease in Primary CNS Lymphoma by Polymerase Chain Reaction of the Rearranged Immunoglobulin Heavy-Chain Genes

Journal of Clinical Oncology ◽

10.1200/jco.2006.06.7165 ◽

2006 ◽

Vol 24 (29) ◽

pp. 4754-4757 ◽

Cited By ~ 55

Author(s):

Kristoph Jahnke ◽

Michael Hummel ◽

Agnieszka Korfel ◽

Thomas Burmeister ◽

Philipp Kiewe ◽

...

Keyword(s):

Bone Marrow ◽

Polymerase Chain Reaction ◽

Systemic Disease ◽

Primary Cns Lymphoma ◽

Cns Lymphoma ◽

Chain Reaction ◽

Blood Samples ◽

Pcr Products ◽

Polymerase Chain

Purpose To search for subclinical systemic disease in bone marrow and peripheral blood in patients with primary CNS lymphoma (PCNSL) to elucidate whether extracerebral relapse may represent a sequel of initial occult systemic disease rather than true extracerebral spread. Patients and Methods Bone marrow and peripheral-blood specimens of 24 PCNSL patients were examined using polymerase chain reaction (PCR) for analysis of clonally rearranged immunoglobulin heavy-chain (IgH) genes. Results Identical dominant PCR products were found in bone marrow aspirates, blood samples, and tumor biopsy specimens of two patients, indicating that the same tumor cell population is present in the CNS and in extracerebral sites. Follow-up IgH PCR performed in one of these patients in complete remission 24 months after diagnosis yielded a persistent monoclonal product in the blood. An oligoclonal IgH rearrangement pattern was found in the tumor specimen of two other patients, whereas bone marrow and blood samples demonstrated the same dominant PCR products. Follow-up PCR showed a persistent monoclonal amplificate in blood in one of these patients 27 months after diagnosis. Conclusion It could be demonstrated for the first time that subclinical systemic disease can be present in PCNSL patients at initial diagnosis. Our findings may have an impact on the understanding of PCNSL pathogenesis and the extent of staging and treatment.

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