3′ RACE LaNe: a simple and rapid fully nested PCR method to determine 3′-terminal cDNA sequence

Park, Daniel J.

doi:10.2144/04364bm04

Cited by 6 publications

(1 citation statement)

References 12 publications

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“…PCR products were cloned using the TOPO TA cloning kit for sequencing (Invitrogen). The 3Ј end of chicken CSF-1 was cloned using a modified 3Ј RACE technique, 3Ј RACE LaNe [26].…”

Section: Cloning Of Chicken and Zebra Finch Cdna Genesmentioning

confidence: 99%

Pivotal Advance: Avian colony-stimulating factor 1 (CSF-1), interleukin-34 (IL-34), and CSF-1 receptor genes and gene products

Garceau

Smith

Paton

et al. 2010

Journal of Leukocyte Biology

154

174

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Macrophages are involved in many aspects of development, host defense, pathology, and homeostasis. Their normal differentiation, proliferation, and survival are controlled by CSF-1 via the activation of the CSF1R. A recently discovered cytokine, IL-34, was shown to bind the same receptor in humans. Chicken is a widely used model organism in developmental biology, but the factors that control avian myelopoiesis have not been identified previously. The CSF-1, IL-34, and CSF1R genes in chicken and zebra finch were identified from respective genomic/cDNA sequence resources. Comparative analysis of the avian CSF1R loci revealed likely orthologs of mammalian macrophage-specific promoters and enhancers, and the CSF1R gene is expressed in the developing chick embryo in a pattern consistent with macrophage-specific expression. Chicken CSF-1 and IL-34 were expressed in HEK293 cells and shown to elicit macrophage growth from chicken BM cells in culture. Comparative sequence and co-evolution analysis across all vertebrates suggests that the two ligands interact with distinct regions of the CSF1R. These studies demonstrate that there are two separate ligands for a functional CSF1R across all vertebrates.

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“…PCR products were cloned using the TOPO TA cloning kit for sequencing (Invitrogen). The 3Ј end of chicken CSF-1 was cloned using a modified 3Ј RACE technique, 3Ј RACE LaNe [26].…”

Section: Cloning Of Chicken and Zebra Finch Cdna Genesmentioning

confidence: 99%

Pivotal Advance: Avian colony-stimulating factor 1 (CSF-1), interleukin-34 (IL-34), and CSF-1 receptor genes and gene products

Garceau

Smith

Paton

et al. 2010

Journal of Leukocyte Biology

154

174

View full text Add to dashboard Cite

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Lariat-Dependent Nested PCR for Flanking Sequence Determination

Park

2010

Methods in Molecular Biology

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Methods detailed in this chapter relate to the use of Lariat-dependent Nested (LaNe) PCR to characterize unknown RNA or DNA sequence flanking known regions. A multitude of approaches designed to determine flanking sequences have been described in the literature. Variously, problems related to these approaches include lack of resolution or failure, depending on experimental context, and complex handling. LaNe-based methods are designed to harness "two-sided" gene-specific PCR with the option of nesting but without the requirement for inefficient and involved enzyme preprocessing steps.

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Complete genomic sequence of an infectious pancreatic necrosis virus isolated from rainbow trout (Oncorhynchus mykiss) in China

et al. 2016

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Infectious pancreatic necrosis (IPN) is a significant disease of farmed salmonids resulting in direct economic losses due to high mortality in China. However, no gene sequence of any Chinese infectious pancreatic necrosis virus (IPNV) isolates was available. In the study, moribund rainbow trout fry samples were collected during an outbreak of IPN in Yunnan province of southwest China in 2013. An IPNV was isolated and tentatively named ChRtm213. We determined the full genome sequence of the IPNV ChRtm213 and compared it with previously identified IPNV sequences worldwide. The sequences of different structural and non-structural protein genes were compared to those of other aquatic birnaviruses sequenced to date. The results indicated that the complete genome sequence of ChRtm213 strain contains a segment A (3099 nucleotides) coding a polyprotein VP2-VP4-VP3, and a segment B (2789 nucleotides) coding a RNA-dependent RNA polymerase VP1. The phylogenetic analyses showed that ChRtm213 strain fell within genogroup 1, serotype A9 (Jasper), having similarities of 96.3% (segment A) and 97.3% (segment B) with the IPNV strain AM98 from Japan. The results suggest that the Chinese IPNV isolate has relative closer relationship with Japanese IPNV strains. The sequence of ChRtm213 was the first gene sequence of IPNV isolates in China. This study provided a robust reference for diagnosis and/or control of IPNV prevalent in China.

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3′ RACE LaNe: a simple and rapid fully nested PCR method to determine 3′-terminal cDNA sequence

Cited by 6 publications

References 12 publications

Pivotal Advance: Avian colony-stimulating factor 1 (CSF-1), interleukin-34 (IL-34), and CSF-1 receptor genes and gene products

Pivotal Advance: Avian colony-stimulating factor 1 (CSF-1), interleukin-34 (IL-34), and CSF-1 receptor genes and gene products

Lariat-Dependent Nested PCR for Flanking Sequence Determination

Complete genomic sequence of an infectious pancreatic necrosis virus isolated from rainbow trout (Oncorhynchus mykiss) in China

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