Cloning of lactoferrin gene of mice and the difference of lactoferrin gene expression in mammary gland at different lactation stages

Total RNA was extracted from mammary gland of mice. Two primers were designed and synthesized according to the murine lactoferrin gene reported in Genbank. A cDNA fragment about 897 bp in length was obtained through RT-PCR. The PCR product was cloned into pGEM-T vector and then sequenced. Sequence a...

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Main Authors: WANG Yi-zhen, HAN Fei-fei, HUANG Hai-qing
Format: Article
Language:English
Published: Zhejiang University Press 2004-01-01
Series:浙江大学学报. 农业与生命科学版
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Online Access:https://www.academax.com/doi/10.3785/1008-9209.2004.01.0083
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author WANG Yi-zhen
HAN Fei-fei
HUANG Hai-qing
author_facet WANG Yi-zhen
HAN Fei-fei
HUANG Hai-qing
author_sort WANG Yi-zhen
collection DOAJ
description Total RNA was extracted from mammary gland of mice. Two primers were designed and synthesized according to the murine lactoferrin gene reported in Genbank. A cDNA fragment about 897 bp in length was obtained through RT-PCR. The PCR product was cloned into pGEM-T vector and then sequenced. Sequence analysis suggested that this fragment corresponded to a portion of the sequence of lactoferrin cDNA and coded 299 amino acid residues, which constituted the major part of mature lactoferrin. The gene homology of fragment obtained in this study was 99% comparable to that of the reported lactoferrin cDNA sequence in uterus of mice. Based on the lactoferrin gene clone, we successfully constructed an optimal semi-quantitative RT-PCR method. Using β-actin as inner control, we detected the difference in expression of lactoferrin gene at different lactation stages of mice. The results showed that the amount of lactoferrin gene expression changed with lactation stages, it decreased from day 1 of lactation to day 18 of lactation, but during involution of mice mammary gland (day 25 of lactation), lactoferrin gene expression increased.
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publishDate 2004-01-01
publisher Zhejiang University Press
record_format Article
series 浙江大学学报. 农业与生命科学版
spelling doaj-art-877bc31dfe2c49dab068cc4091af62ac2025-08-20T03:58:14ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552004-01-0130838810.3785/1008-9209.2004.01.008310089209Cloning of lactoferrin gene of mice and the difference of lactoferrin gene expression in mammary gland at different lactation stagesWANG Yi-zhenHAN Fei-feiHUANG Hai-qingTotal RNA was extracted from mammary gland of mice. Two primers were designed and synthesized according to the murine lactoferrin gene reported in Genbank. A cDNA fragment about 897 bp in length was obtained through RT-PCR. The PCR product was cloned into pGEM-T vector and then sequenced. Sequence analysis suggested that this fragment corresponded to a portion of the sequence of lactoferrin cDNA and coded 299 amino acid residues, which constituted the major part of mature lactoferrin. The gene homology of fragment obtained in this study was 99% comparable to that of the reported lactoferrin cDNA sequence in uterus of mice. Based on the lactoferrin gene clone, we successfully constructed an optimal semi-quantitative RT-PCR method. Using β-actin as inner control, we detected the difference in expression of lactoferrin gene at different lactation stages of mice. The results showed that the amount of lactoferrin gene expression changed with lactation stages, it decreased from day 1 of lactation to day 18 of lactation, but during involution of mice mammary gland (day 25 of lactation), lactoferrin gene expression increased.https://www.academax.com/doi/10.3785/1008-9209.2004.01.0083lactoferrinmicemammary glandgene clonesemi-quantitative RT-PCRgene expression
spellingShingle WANG Yi-zhen
HAN Fei-fei
HUANG Hai-qing
Cloning of lactoferrin gene of mice and the difference of lactoferrin gene expression in mammary gland at different lactation stages
浙江大学学报. 农业与生命科学版
lactoferrin
mice
mammary gland
gene clone
semi-quantitative RT-PCR
gene expression
title Cloning of lactoferrin gene of mice and the difference of lactoferrin gene expression in mammary gland at different lactation stages
title_full Cloning of lactoferrin gene of mice and the difference of lactoferrin gene expression in mammary gland at different lactation stages
title_fullStr Cloning of lactoferrin gene of mice and the difference of lactoferrin gene expression in mammary gland at different lactation stages
title_full_unstemmed Cloning of lactoferrin gene of mice and the difference of lactoferrin gene expression in mammary gland at different lactation stages
title_short Cloning of lactoferrin gene of mice and the difference of lactoferrin gene expression in mammary gland at different lactation stages
title_sort cloning of lactoferrin gene of mice and the difference of lactoferrin gene expression in mammary gland at different lactation stages
topic lactoferrin
mice
mammary gland
gene clone
semi-quantitative RT-PCR
gene expression
url https://www.academax.com/doi/10.3785/1008-9209.2004.01.0083
work_keys_str_mv AT wangyizhen cloningoflactoferringeneofmiceandthedifferenceoflactoferringeneexpressioninmammaryglandatdifferentlactationstages
AT hanfeifei cloningoflactoferringeneofmiceandthedifferenceoflactoferringeneexpressioninmammaryglandatdifferentlactationstages
AT huanghaiqing cloningoflactoferringeneofmiceandthedifferenceoflactoferringeneexpressioninmammaryglandatdifferentlactationstages