To construct an expression vector of human anti-keratin scFv and express functional scFv in E. coli.The V(H) and V(L) genes were amplified by PCR and cloned into a scFv expression vector, which was expressed in E. coli. The expression of scFv was tested by SDS-PAGE; The antigen-binding characteristics of scFv were tested by ELISA.The V(H) and V(L) genes were cloned and expressed in E. coli successfully. The expressed scFv retained the antigen-binding characteristics of the parental antibody.A functional anti-keratin scFv was obtained.
Pre-implantation embryo metabolism demonstrates distinctive characteristics associated with the development potential of embryos. We aim to determine if metabolic differences correlate with embryo morphology. In this study, gas chromatography - mass spectroscopy (GC-MS)-based metabolomics was used to assess the culture media of goat cloned embryos collected from high-quality (HQ) and low-quality (LQ) groups based on morphology. Expression levels of amino acid transport genes were further examined by quantitative real-time PCR. Results showed that the HQ group presented higher percentages of blastocysts compared with the LQ counterparts (P < 0.05). Metabolic differences were also present between HQ and LQ groups. The culture media of the HQ group showed lower levels of valin, lysine, glutamine, mannose and acetol, and higher levels of glucose, phytosphingosine and phosphate than those of the LQ group. Additionally, expression levels of amino acid transport genes SLC1A5 and SLC3A2 were significantly lower in the HQ group than the LQ group (P < 0.05, respectively). To our knowledge, this is the first report which uses GC-MS to detect metabolic differences in goat cloned embryo culture media. The biochemical profiles may help to select the most in vitro viable embryos.
Reproductive ability, especially prolificacy, impacts sheep profitability. Hu sheep, a unique Chinese breed, is recognized for its high prolificacy (HP), early sexual maturity, and year-round estrus. However, little is known about the molecular mechanisms underlying HP in Hu sheep. To explore the potential mRNAs and long non-coding RNAs (lncRNAs) involved in Hu sheep prolificacy, we performed an ovarian genome-wide analysis of mRNAs and lncRNAs during the follicular stage using Hu sheep of HP (litter size = 3; three consecutive lambings) and low prolificacy (LP, litter size = 1; three consecutive lambings). Plasma luteinizing hormone (LH) concentration was higher in the HP group than in the LP group (P<0.05) during the follicular stage. Subsequently, 76 differentially expressed mRNAs (DE-mRNAs) and five differentially expressed lncRNAs (DE-lncRNAs) were identified by pairwise comparison; quantitative real-time PCR (qRT-PCR) analysis of ten randomly selected DE genes (mRNA and lncRNA) were consistent with the sequencing results. Gene Ontology (GO) analysis of DE-mRNAs revealed significant enrichment in immune response components, actin filament severing and phagocytosis. Pathway enrichment analysis of DE-mRNAs indicated a predominance of immune function pathways, including phagosomes, lysosomes, and antigen processing. We constructed a co-expression network of DE-mRNAs and mRNA-lncRNAs, with C1qA, CD53, cathepsin B (CTSB), CTSS, TYROBP, and AIF1 as the hub genes. Finally, the expression of lysosomal protease cathepsin genes, CTSB and cathepsin D (CTSD), were significantly up-regulated in sheep ovaries in the HP group compared with the LP group (P<0.05). These differential mRNAs and lncRNAs may provide information on the molecular mechanisms underlying sheep prolificacy.
Chickens and ducks were raised for controlling locusts in the high and cold pastureland of Qinghai Lake.The results showed that the elimination rate reached up to 91.49% during the 60 days-experiment.It provided the base of application the natural enemy to control insects in high and cold pastureland.
The DNA sequence recognition study of DNA-targeted anticancer drugs is a theoretical basis for improving the selectivity of anticancer drugs. With the high synergy effect of cocoamidopropyl hydroxy sulfobetaine (HSB), a resonance light scattering (RLS) quenching system for DNA sequence recognition studies of actinomycin D (ACTD) was developed in this contribution. By the strategy, DNA sequence selectivity as well as the recognition mechanisms of ACTD was systematically investigated. The results suggested that ACTD had the selectivity to single-stranded DNA (ssDNA) with an equilibrium constant (K(RLS)) of 12.4 mmol mg(-1). Also it had a preference for Guanine and Cytosine bases with a K(RLS) of 6.69 L mmol(-1). The selectivity mechanism between ACTD and DNA was also well discussed with the help of UV-Vis absorption spectroscopy. Compared with other methods, the RLS quenching system has the advantages of reliability and speediness, and it avoids complex modification processes and is a better bionic system for the above research. Results obtained from this work would supply a theoretical basis for improving anticancer activity and designing similar anticancer drugs.
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