Quantification of silkworm coactivator of MBF1 mRNA by SYBR Green I real-time RT-PCR reveals tissue- and stage-specific transcription levels.
Transcriptional coactivators play a crucial role in gene transcription and expression. Multiprotein bridging factor 1 (MBF1) is a transcriptional coactivator necessary for transcriptional activation caused by DNA-binding activators, such as FTZ-F1 and GCN4. Until now, very few studies have been reported in the silkworm. We selected the Bombyx mori because it is a model insect and acts as an economic animal for silk industry. In this study, we conducted the quantitative analysis of MBF1 mRNA in silkworm B. mori L. with actin (A3) as internal standard by means of SYBR Green I real-time RT-PCR method. The total RNA was extracted from the silk gland, epidermis, fat body, and midguts of the fifth instar B. mori larvae. The mRNA was reverse transcripted, and the cDNA fragments of MBF1 mRNA and actin gene were amplified by RT-PCR using specific primers. MBF1 mRNA expression in different tissues of silkworm B. mori L. was quantified using standardized SYBR Green I RT-PCR. The results suggested MBF1 gene was expressed in all investigated organs but highly expressed in the silk gland, showing its relation to biosynthesis of silk proteins.
Li
Guang-li
GL
College of Animal Sciences, Institute of Sericulture and Apiculture, Zhejiang University, Hangzhou, People's Republic of China.
Roy
Bhaskar
B
Li
Xing-hua
XH
Yue
Wan-fu
WF
Wu
Xiao-feng
XF
Liu
Jian-mei
JM
Zhang
Chuan-xi
CX
Miao
Yun-gen
YG
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D052160:Benzothiazoles; D012831:Bombyx / Q000235:genetics* / Q000254:growth & development*; D003001:Cloning, Molecular; D003959:Diamines; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics*; D008018:Life Cycle Stages / Q000235:genetics*; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics*; D009930:Organic Chemicals / Q000378:metabolism; D011804:Quinolines; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
D052160:Benzothiazoles; D003959:Diamines; D019476:Insect Proteins; D009930:Organic Chemicals; D011804:Quinolines; D012333:RNA, Messenger; C098022:SYBR Green I