RESUMEN
Sinopodophyllum hexandrum is an important medicinal plant that has been listed as an endangered species, making the conservation of its genetic diversity a priority. Therefore, the genetic diversity and population structure of S. hexandrum was investigated through inter-simple sequence repeat analysis of eight natural populations. Eleven selected primers generated 141 discernible fragments. The percentage of polymorphic bands was 37.59% at the species level, and 7.66-24.32% at the population level. Genetic diversity of S. hexandrum was low within populations (average HE = 0.0366), but higher at the species level (HE = 0.0963). Clear structure and high genetic differentiation were detected between populations using unweighted pair groups mean arithmetic and principle coordinate analysis. Clustering approaches clustered the eight sampled populations into three major groups, and AMOVA confirmed there to be significant variation between populations (63.27%). Genetic differentiation may have arisen through limited gene flow (Nm = 0.3317) in this species. Isolation by distance among populations was determined by comparing genetic distance versus geographical distance using the Mantel test. The results revealed no correlation between spatial pattern and geographic location. Given the low within-population genetic diversity, high differentiation among populations, and the increasing anthropogenic pressure on this species, in situ conservation measures, in addition to sampling and ex situ preservation, are recommended to preserve S. hexandrum populations and to retain their genetic diversity.
Asunto(s)
Berberidaceae/genética , Variación Genética , Repeticiones de Microsatélite/genética , Filogeografía , Berberidaceae/crecimiento & desarrollo , Conservación de los Recursos Naturales , Especies en Peligro de Extinción , Flujo Genético , Genética de PoblaciónRESUMEN
An antifungal protein exhibiting a high activity against Sclerotinia sclerotiorum in vivo was purified by ammonium sulfate precipitation, hydrophobic chromatography, and gel filtration chromatography from the culture filtrate of the endophytic Bacillus subtilis strain Em7. The protein was characterized as a ß-1,3-1,4-glucanase according to amino acid analysis, and showed excellent properties in thermal stability and acid resistance. At the same time, the antifungal protein was cloned and heterologously expressed in Escherichia coli BL21. The recombinant protein was purified and showed similar enzymatic properties to the native protein, exhibiting strong inhibitory activity against S. sclerotiorum. This shows that the ß-1,3-1,4-glucanase may play a very important role in B. subtilis Em7 biocontrol function. In addition, many physiochemical properties of the native and purified recombinant protein were compared, including the effect of pH, temperature, metal cations, substrate specificity, and kinetic parameters. All parameters were similar between the native and recombinant purified protein, indicating that the purified recombinant protein has potential for industrial applications.
Asunto(s)
Antifúngicos/farmacología , Ascomicetos/efectos de los fármacos , Bacillus subtilis/genética , Proteínas Bacterianas/genética , Proteínas Bacterianas/farmacología , Proteínas Recombinantes , Proteínas Bacterianas/química , Proteínas Bacterianas/aislamiento & purificación , Activación Enzimática , Escherichia coli/genética , Escherichia coli/metabolismo , Expresión Génica , Cinética , Pruebas de Sensibilidad Microbiana , Estabilidad Proteica , Especificidad por SustratoRESUMEN
Acute ischemic stroke (AIS) has become a serious health problem in many countries because of its poor outcome and worsening epidemic trend. Early identification of genetic risk factors and physiological indicators for stroke occurrence may help to reduce the incidence of stroke. Therefore, we conducted a case-control study including 50 AIS patients and 50 healthy individuals from a Chinese population to explore the association between AIS and patient complete blood profiles and the association between AIS and the genetic polymorphism K469E in intercellular adhesion molecule-1 (ICAM-1). Compared to the control group, AIS patients showed a high percentage of mononuclear cells, low platelet count, low ratio of platelet to lymphocyte count, high frequency of the 469K allele, and low frequency of the 469E allele. White blood cell count, percentage of neutrophils, percentage of lymphatic cells, platelet distribution width, mean platelet volume, and platelet hematocrit levels showed no significant differences between the 2 groups and between different genotypes. Our results suggested an association of elevated levels of mononuclear cells and reduced platelet count with higher AIS risk. Our results also supported the hypothesis that the KK genotype at the K469E locus in ICAM-1 is a risk factor for AIS.
Asunto(s)
Predisposición Genética a la Enfermedad , Molécula 1 de Adhesión Intercelular/genética , Fenotipo , Polimorfismo de Nucleótido Simple , Accidente Cerebrovascular/etiología , Accidente Cerebrovascular/patología , Adulto , Anciano , Anciano de 80 o más Años , Alelos , Sustitución de Aminoácidos , Estudios de Casos y Controles , Índices de Eritrocitos , Femenino , Frecuencia de los Genes , Genotipo , Pruebas Hematológicas , Humanos , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Recuento de Plaquetas , Análisis de Secuencia de ADN , Accidente Cerebrovascular/sangre , Adulto JovenRESUMEN
Geese are an economically important poultry species worldwide. Their superior meat production performance and meat qual-ity make them a popular food. However, they are not bred worldwide because their poor laying capacity increases farming costs. To gain a global view of the genes that are differentially expressed between pre-laying (P) and laying (L) periods and to develop a database for further studies, we performed large-scale transcriptome sequencing of ovarian tissue collected from Anser cygnoides. In total, 30,151,422 raw reads, with an average length of 151 bp and a total length of 4,552,864,722 bp, were obtained. After primers and adaptors were removed, 19,167,132 clean reads, with an average length of 134.5 bp and a total length of 2,577,297,281 bp, were obtained, among which 1,268,906,694 bp and 1,308,390,587 bp were from L and P ovarian tissue, respectively. The 16,605 assembled sequences were further functionally annotated by comparing their sequences to different protein and functional domain databases and assigning gene ontology (GO) terms. Of these, 511 as-sembled sequences were considered differentially expressed based on the 2-fold method, among which 396 were assigned at least one GO term. Digital expression analysis using the Kyoto encyclopedia of genes and genomes annotation identified 121 genes that were differ-entially expressed in the P vs L periods. Five of these are of special interest for further investigation of their roles in determining high re-productive performance. This study provides valuable information and sequence resources for uncovering genes determining high egg-laying performance and for future functional genomics analysis of geese.
Asunto(s)
Gansos/genética , Regulación de la Expresión Génica , Aptitud Genética/genética , Genoma , Oviposición/genética , Transcriptoma , Animales , Mapeo Cromosómico , Femenino , Perfilación de la Expresión Génica , Anotación de Secuencia Molecular , Ovario/metabolismo , Carácter Cuantitativo HeredableRESUMEN
PURPOSE: The response rate of first-line fluoropyrimidine-based regimens for metastatic colorectal cancer (mCRC) is generally less than 50 %. The down-regulation of miR-197 in colorectal cancer cells after exposure to 5-fluorouracil might be related to the mechanism of resistance to fluoropyrimidine-based chemotherapy. So we investigated the regulatory mechanism of miR-197 on 5-FU sensitivity. METHODS: Dual luciferase reporter gene construct and dual luciferase reporter assay were used to identify the target of miR-197. TYMS expression was evaluated by immunohistochemistry staining. 5-Fu resistance of colorectal cancer cell lines was detected by MTS assay. The expression of miR-197 was detected by real time PCR. RESULTS: A luciferase assay and western blot analysis confirmed that miR-197 directly binds to and negatively regulates TYMS expression. Overexpressing miR-197 could increase the sensitivity of colorectal cancer cells to 5-fluorouracil (5-FU). The expression of miR-197 negatively correlated with TYMS expression in cancerous tissues from patients with stage IV colorectal cancer. CONCLUSION: miR-197 mediates the response of colorectal cancer cells to 5-FU by regulating TYMS expression.
Asunto(s)
Neoplasias Colorrectales/genética , Resistencia a Antineoplásicos/genética , Regulación Neoplásica de la Expresión Génica/genética , MicroARNs/genética , Timidilato Sintasa/biosíntesis , Antimetabolitos Antineoplásicos/uso terapéutico , Western Blotting , Línea Celular Tumoral , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/enzimología , Fluorouracilo/uso terapéutico , Humanos , Inmunohistoquímica , ARN Interferente Pequeño , Reacción en Cadena en Tiempo Real de la Polimerasa , TransfecciónRESUMEN
A mouse model of acute lung injury (ALI) was chosen in this study to explore the key genes and pathways involved in the process of ALI with microarray technology. Gene expression microarray data were downloaded from the Gene Expression Omnibus database. Mice from the experimental group were further divided into 6 subgroups, which received octadecenoate treatments for 1, 1.5, 3, 4, 18, and 24 h. Differentially co-expressed genes were screened to uncover the pathogenesis of ALI. Almost all of the differentially co-expressed genes were identified at two times: 1.5 and 3 h. Functional analysis revealed that several inflammation-related pathways were significantly enriched. Ubiquitin-mediated proteolysis, hematopoietic cell lineage, and leukocyte transendothelial migration were enriched at 1.5 h. The B cell receptor signaling pathway, T cell receptor signaling pathway, natural killer cell-mediated cytotoxicity, Fc epsilon RI signaling pathway, and ubiquitin-mediated proteolysis were significantly enriched at 3 h. It could be inferred that ALI initiated at 1.5 h and lasted through 3 h. However, co-expression patterns were not found from 4 h onward. In conclusion, several key genes and pathways implicated in the development of ALI were found in this study using the mouse model, among which ubiquitin-mediated proteolysis appears to play an important role in the process.