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Establishment and Application of Target Gene Disruption System in Saccharomyces boulardii
Longjiang Wang,Hui Sun,Jie Zhang,Qing Liu,Tiantian Wang,Peipei Chen,Hongmei Li,Yihong Xiao,Fangkun Wang,Xiaomin Zhao 한국생물공학회 2015 Biotechnology and Bioprocess Engineering Vol.20 No.1
Saccharomyces boulardii is the best knownprobiotic yeast, widely used as a therapeutic agent for thetreatment or prevention of diarrhea and intestine disorders. In the present work, we established a target gene disruptionsystem for S. boulardii based on the Cre-loxP system usedfor S. cerevisiae and other fungi by screening out selectionmarkers, working out the transformation method, andconstructing essential plasmids for S. boulardii. Theestablished system was successfully applied to the URA3gene disruption and created an ura3 null mutant strain ofS. boulardii. The system can be used for PCR mediatedgene disruption, cloning mediated gene disruption, andreintroduction of the deleted gene back to the mutant. Allthe introduced exogenous DNAs in the gene disruptionprocedures were removed from the final mutant strainexcept the two 34 bp loxP pieces left in deleted gene loci.
Li Maoteng,Xiang Jun,Liu Jianmin,Yu Longjiang,Li Dianrong 한국유전학회 2008 Genes & Genomics Vol.30 No.2
The Brassica napus-B genome monosomic addition lines (MALs) (AACC + B’, 2n = 39) were developed from self-pollination of pentaploid hybrids (AABCC) that were derived from hybridization between hexaploid hybrids (AABBCC) and B. napus (AACC). The alien chromosomes of the B genome in MALs were identified by the GISH technique, by observation of the meiotic behavior of pollen mother cells (PMCs), and by B-genome-specific molecular marker analysis. Studies of the meiotic behavior of B. napus-B genome chromosome MALs at diakinesis revealed that the majority of the chromosome configuration was 19²+1¹, which indicated that the alien B genome chromosome remained univalent in most cases. The laggard-free PMCs also appeared at a lower ratio, which indicated that the B genome chromosome could be transmitted into gametes. The chromosome configurations of 20² and 19²+2¹ that appeared in double MALs (AACC+ 2 chromosomes of the B genome) indicated different homoeology between different B genome chromosomes. The paired B genome bivalent in double MALs can be normally segregated at anaphase in most cases. PMCs with multivalents were observed in all the double MAL combinations, which indicated homology of the B genome chromosomes with the A or C genome chromosomes. The Brassica napus-B genome monosomic addition lines (MALs) (AACC + B’, 2n = 39) were developed from self-pollination of pentaploid hybrids (AABCC) that were derived from hybridization between hexaploid hybrids (AABBCC) and B. napus (AACC). The alien chromosomes of the B genome in MALs were identified by the GISH technique, by observation of the meiotic behavior of pollen mother cells (PMCs), and by B-genome-specific molecular marker analysis. Studies of the meiotic behavior of B. napus-B genome chromosome MALs at diakinesis revealed that the majority of the chromosome configuration was 19²+1¹, which indicated that the alien B genome chromosome remained univalent in most cases. The laggard-free PMCs also appeared at a lower ratio, which indicated that the B genome chromosome could be transmitted into gametes. The chromosome configurations of 20² and 19²+2¹ that appeared in double MALs (AACC+ 2 chromosomes of the B genome) indicated different homoeology between different B genome chromosomes. The paired B genome bivalent in double MALs can be normally segregated at anaphase in most cases. PMCs with multivalents were observed in all the double MAL combinations, which indicated homology of the B genome chromosomes with the A or C genome chromosomes.
TCP10L negatively regulates alpha-fetoprotein expression in hepatocellular carcinoma
Suqin Shen,Huan Feng,Longjiang Liu,Wei Su,Jiaxue Wu,Long Yu 생화학분자생물학회 2020 BMB Reports Vol.53 No.8
Alpha-fetoprotein (AFP) is one of the most commonly used and reliable biomarkers for Hepatocellular carcinoma (HCC). However, the underlying mechanism of AFP expression in HCC is poorly understood. In this study, we found that TCP10L, a gene specifically expressed in the liver, is down-regulated in HCC and that its expression inversely correlates with AFP expression. Moreover, overexpression of TCP10L suppresses AFP expression whereas knockdown of TCP10L increases AFP expression, suggesting that TCP10L might be a negative regulator of AFP. We found that TCP10L is associated with the AFP promoter and inhibits AFP promoter-driven transcriptional activity. Taken together, these results indicate that TCP10L negatively regulates AFP expression in HCC and that it could be a potential prognostic marker and therapeutic target for HCC.