Characterization of a potato Proteinase Inhibitor II gene whose expression in not wound-responsive

A potato Proteinase Inhibitor II gene (pin2T) was isolated and characterized both at the molecular level and the functional level. The open reading frame as well as the 5[superscript]\u27 and 3[superscript]\u27 flanking regions were sequenced. The coding region and 3[superscript]\u27 flanking region showed 86% and 87% identity with those of the previously isolated wound-inducible potato Proteinase Inhibitor IIK gene (pin2K) sequence, respectively. However, the 5[superscript]\u27 flanking region of the pin2T is highly homologous (91% identity) with that of pin2K from -767 to +29, relative to the transcription start site of the wound-inducible pin2K. The 5[superscript]\u27 flanking region of pin2T was linked to the reporter gene (chloramphenicol acetyl transferase, CAT; or [beta]-glucuronidase, GUS) coding sequences in constructions that contained the terminator from the wound-inducible pin2K gene. The chimeric genes were transferred to tobacco plants using Agrobacterium tumefaciens. The presence of the constructions in the transgenic tobacco plants was confirmed by polymerase chain reaction (PCR). Expression of CAT or GUS activities in transgenic plants driven by pin2T promoter indicated that pin2T is not a wound-inducible gene. GUS activities driven by promoter deletion mutants ruled out the presence of the silencer sequences in upstream portion of the pin2T promoter that differed from pin2K. Comparison of pin2T promoter sequence with that of the wound-inducible pin2K indicates that there are four small deletions which are located at -221 to -200, -263 to -254, -523 to -462, and -759 to -708 relative to the transcription start site of pin2K. When these deleted sequences are searched through Genebank, we identified one sequence that is found three times in pin2K and is completely deleted from pin2T. This sequence, 5[superscript]\u27-AGTAAA-3[superscript]\u27, is found in a wide variety of other wound-inducible genes but is not easily found in the published promoter sequences of the other plant genes. Nuclear proteins from unwounded and wounded potato leaves bound to the proximal promoter region, downstream of the 5[superscript]\u27-AGTAAA-3[superscript]\u27, of pin2T. A third deletion located at -523 to -462 relative to the transcription initiation site of pin2K, was moderately homologous (72% identity) with a putative sucrose-responsive element of sucrose-inducible genes. Deletion of this sequence was correlated with a loss of sucrose inducibility in the pin2T promoter. CAT activity driven by pin2T was not enhanced by sucrose, while sucrose enhances the expression of pin2K-CAT construct in transgenic tobacco plants. In addition to the sucrose effect on the chimeric gene expression, the effects of plant hormones, abscisic acid (ABA) and [alpha]-naphthalene acetic acid ([alpha]-NAA), were examined in transgenic tobacco leaves and calli, respectively. CAT activity driven by the non-wound-inducible pin2T promoter was not induced by ABA or derepressed in the absence of [alpha]-NAA as was the wound-inducible pin2K promoter

A C/C++-based functional verification framework using the SystemC verification library_

This paper describes SoCBase-VL, which is a C/C++ based integrated framework for SoC functional verification. It has a layered architecture which provides easier testbench description, automatic verification of bus interfaces and seamless testbench migration. This framework does not require verification engineers to learn other verification languages as long as they have sufficient knowledge on both C/C++ and SystemC. We have confirmed its usefulness by applying it to a TFT-LCD Controller verification

Reusable Component IP Design using Refinement-based Design Environment

We propose a method of enhancing the reusability of the component IPs by separating communication and computation for a system function. In this approach, we assume that the component designers describe mainly the computation part of the component, and the system designer can construct the communication part by using our refinement-based design environment. Moreover, we introduced a concept of the Communication Architecture Template Tree (CATree), which helps IP designers to effectively separate computation and communication for a system function. We confirmed that this approach is effective by applying it to a H.264 decoder design

Implementation of a H.264 decoder with Template-based Communication Refinement

We described an H.264 decoder implemented with our design methodology, in which a system function model of transaction level is first captured in SystemC and refined into RTL with a library of communication templates. We determined its communication architecture by exploring the design space with template-based communication refinement to meet its requirement of decoding VGA 30 frames per second at a clock frequency of 50MHz

A mixed-level virtual prototyping environment for refinement-based design environment

The Communication Architecture Template Tree (CATtree) is an abstraction of the specific range of communication functions and architectures, which can facilitate system function capture and communication architecture refinement. In this paper, we explain a TLM-RTL-SW mixedlevel simulation environment that is useful for the functional verification of partially refined system models. We employed SystemC, GNU Gdb and a HDL simulator for the simulation of CATtree-based TLM, SW and HW, respectively. We also employed a new operating system, DEOS so that each SystemC-based TLMs can be cross-compiled to be executed as software models on the target processors. We evaluated the flexibility and simulation performance of the virtual simulation environment with an H.264 decoder design example

Introduction of Transmembrane Inner Ear (tmie) Gene Can Recover the Hearing Impairment and Abnormal Behavior in the Circling Mouse

The spontaneous mutant circling mouse (cir/cir) shows a circling behavior and hearing loss. We produced transgenic mice overexpressing the causative gene, transmembrane inner ear (tmie), for the phenotypic rescue of the circling mouse. Through the continuous breeding with circling mice, the cir/cir homozygous mice carrying the transgene (cir/cir-tg) were produced. The rescued cir/cir -tg mice were able to swim in the water with proper orientation and did not show any circling behavior like wild type mice. Western blot and immunohistochemical analysis exhibited that the transgenic tmie was expressed in the inner ear. Inner and outer hair cells were recovered in the cochlea and spiral ganglion neurons were also recovered in the rescued mice. Auditory brainstem response (ABR) test demonstrated that the cir/cir -tg mice are able to respond to sound. This study demonstrates that tmie transgene can recover the hearing impairment and abnormal behavior in the circling mouse

JAZF1 heterozygous knockout mice show altered adipose development and metabolism

Background: Juxtaposed with another zinc finger protein 1 (JAZF1) is associated with metabolic disorders, including type 2 diabetes mellitus (T2DM). Several studies showed that JAZF1 and body fat mass are closely related. We attempted to elucidate the JAZF1 functions on adipose development and related metabolism using in vitro and in vivo models. Results: The JAZF1 expression was precisely regulated during adipocyte differentiation of 3T3-L1 preadipocyte and mouse embryonic fibroblasts (MEFs). Homozygous JAZF1 deletion (JAZF1-KO) resulted in impaired adipocyte differentiation in MEF. The JAZF1 role in adipocyte differentiation was demonstrated by the regulation of PPARγ—a key regulator of adipocyte differentiation. Heterozygous JAZF1 deletion (JAZF1-Het) mice fed a normal diet (ND) or a high-fat diet (HFD) had less adipose tissue mass and impaired glucose homeostasis than the control (JAZF1-Cont) mice. However, other metabolic organs, such as brown adipose tissue and liver, were negligible effect on JAZF1 deficiency. Conclusion: Our findings emphasized the JAZF1 role in adipocyte differentiation and related metabolism through the heterozygous knockout mice. This study provides new insights into the JAZF1 function in adipose development and metabolism, informing strategies for treating obesity and related metabolic disorders. © 2021, The Author(s).1

Characterization of a potato Proteinase Inhibitor II gene whose expression in not wound-responsive

A potato Proteinase Inhibitor II gene (pin2T) was isolated and characterized both at the molecular level and the functional level. The open reading frame as well as the 5[superscript]' and 3[superscript]' flanking regions were sequenced. The coding region and 3[superscript]' flanking region showed 86% and 87% identity with those of the previously isolated wound-inducible potato Proteinase Inhibitor IIK gene (pin2K) sequence, respectively. However, the 5[superscript]' flanking region of the pin2T is highly homologous (91% identity) with that of pin2K from -767 to +29, relative to the transcription start site of the wound-inducible pin2K. The 5[superscript]' flanking region of pin2T was linked to the reporter gene (chloramphenicol acetyl transferase, CAT; or [beta]-glucuronidase, GUS) coding sequences in constructions that contained the terminator from the wound-inducible pin2K gene. The chimeric genes were transferred to tobacco plants using Agrobacterium tumefaciens. The presence of the constructions in the transgenic tobacco plants was confirmed by polymerase chain reaction (PCR). Expression of CAT or GUS activities in transgenic plants driven by pin2T promoter indicated that pin2T is not a wound-inducible gene. GUS activities driven by promoter deletion mutants ruled out the presence of the silencer sequences in upstream portion of the pin2T promoter that differed from pin2K. Comparison of pin2T promoter sequence with that of the wound-inducible pin2K indicates that there are four small deletions which are located at -221 to -200, -263 to -254, -523 to -462, and -759 to -708 relative to the transcription start site of pin2K. When these deleted sequences are searched through Genebank, we identified one sequence that is found three times in pin2K and is completely deleted from pin2T. This sequence, 5[superscript]'-AGTAAA-3[superscript]', is found in a wide variety of other wound-inducible genes but is not easily found in the published promoter sequences of the other plant genes. Nuclear proteins from unwounded and wounded potato leaves bound to the proximal promoter region, downstream of the 5[superscript]'-AGTAAA-3[superscript]', of pin2T. A third deletion located at -523 to -462 relative to the transcription initiation site of pin2K, was moderately homologous (72% identity) with a putative sucrose-responsive element of sucrose-inducible genes. Deletion of this sequence was correlated with a loss of sucrose inducibility in the pin2T promoter. CAT activity driven by pin2T was not enhanced by sucrose, while sucrose enhances the expression of pin2K-CAT construct in transgenic tobacco plants. In addition to the sucrose effect on the chimeric gene expression, the effects of plant hormones, abscisic acid (ABA) and [alpha]-naphthalene acetic acid ([alpha]-NAA), were examined in transgenic tobacco leaves and calli, respectively. CAT activity driven by the non-wound-inducible pin2T promoter was not induced by ABA or derepressed in the absence of [alpha]-NAA as was the wound-inducible pin2K promoter.</p

A two-week program for platform-based SoC design

This paper describes a two-week program for a platform-based SoC design using SoCBase 1.0, a platform developed in the Center for SoC Design Technology. This program consists of 4 lectures and 9 labs. It covers several design steps from the transaction level to the FPGA prototype level for a Motion JPEG decoder. In this program we employed an SoC design flow based on SoCBase 1.0. It is targeted for graduate students and ASIC designers out in the industry. More than 100 engineers and graduate students have completed this program in 2004

A C/C++-based Functional Verification Framework using the SystemC Verification Library

Abstrac