Cloning and Expression Pattern Studies of a Novel Gene – N4bp2l2

• Main Authors: Yun-Yu Liao, 廖芸郁
• Other Authors: Hu-Hui Lee
• Format: Others
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/69540352750755484288

碩士 === 國立嘉義大學 === 生物農業科技學系碩士班 === 100 === NEDD4 binding protein 2-like 2 (N4bp2l2) is a newly discovered gene which is also termed as phosphonoformate immunoassociated protein 5 (PFAAP5) and zygotic activating gene 1 (ZAG1) based on its distinct functions elicited by different groups. Functional analysis indicated that PFAAP5 can be a feedback regulator to affect neutrophil development, and ZAG1 regulates the normal zygotic proliferation. As for the expression of N4bp2l2, it is repressed by the Osr2 activity that is necessary for osteogenesis. Also Osr2 is regulated by the BMP signaling that plays a major role in osteogenesis. Altogether, N4bp2l2 may function in many aspects including osteoblast differentiation. To further realize the function of N4bp2l2, I examined its expression pattern in the beginning. Northern blot and semi-quantitative RT-PCR were performed in many tissues, showing that N4bp2l2 is expressed ubiquitously, yet most abundant in testis, liver and thymus. During the differentiation of MC3T3-E1, a pre-osteoblast cell, I further examined the expression of N4bp2l2 and marker genes by semi-quantitative RT-PCR in a fixed time interval. I found that the expressions of N4bp2l2 as well as the marker genes were abruptly elevated after 8 day of differentiation. This implies the possible role of N4bp2l2 in osteogenesis. In addition to the full length of N4bp2l2, two more N4bp2l2 isoforms were also identified. Two isoforms are generated due to alternative splicing, which results in lacking of either exon 4 or exon 4+ 5. I immunostained one of the isoforms, and confirmed whose expression is localized within the nucleus as the full length of N4bp2l2. Since the sequences of full length N4bp2l2 comprises of several conserved domains that implies the possible multiple-function of N4bp2l2, and need to be further studied.